==================================BSR10================================== 10. Human adenocarcinoma cell lines, or primary tissue culture cell lines from human adenocarcinomas. 1 UI - 87119118 AU - Kelland LR ; Steel GG TI - Dose-rate effects in the radiation response of four human tumour xenografts. AB - The radiation repair capability of four human tumour xenograft lines, two adenocarcinomas of the pancreas, an adenocarcinoma of the breast and a melanoma, has been investigated by means of a soft agar clonogenic assay. Dose-rate dependence studies at 150, 7.6 and 1.6 cGy/min and split-dose experiments, have been performed. Results indicate a good correlation between split-dose recovery and the dose-sparing achieved by irradiation at 1.6 cGy/min when compared to acute irradiation in 3 of the 4 tumour lines. The HX118 melanoma showed dose-sparing at dose-rates of 7.6 and 1.6 cGy/min and a substantial ability to perform recovery between split doses of radiation, whereas HX32, a pancreatic carcinoma, showed little dose-sparing and a correspondingly small degree of split-dose recovery. The HX99 breast carcinoma, was aberrant in showing the greatest split-dose recovery of the four lines but only a moderate extent of low dose-rate sparing. Data have been fitted by two recently described models for radiation dose-rate dependence; the incomplete repair model of Thames and the Lethal Potentially-Lethal (LPL) model of Curtis. Curves were fitted better by the Thames model which provided a good fit for data from 3 of the 4 xenografted human tumour lines. HX99 produced dose-rate dependence survival curves that were not well fitted by either model. MH - Adenocarcinoma/*RADIOTHERAPY ; Animal ; Breast Neoplasms/*RADIOTHERAPY ; Cell Line ; Cell Survival/*RADIATION EFFECTS ; Cobalt Radioisotopes/ THERAPEUTIC USE ; *Colony-Forming Units Assay ; Female ; Human ; Male ; Melanoma/*RADIOTHERAPY ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Pancreatic Neoplasms/*RADIOTHERAPY ; Radioisotope Teletherapy ; Radiotherapy Dosage ; Skin Neoplasms/*RADIOTHERAPY ; Support, U.S. Gov't, P.H.S. ; *Tumor Stem Cell Assay SO - Radiother Oncol 1986 Nov;7(3):259-68 2 UI - 87113988 AU - Holinka CF ; Hata H ; Gravanis A ; Kuramoto H ; Gurpide E TI - Effects of estradiol on proliferation of endometrial adenocarcinoma cells (Ishikawa line). AB - Estradiol (E2) stimulates the proliferation of human endometrial adenocarcinoma cells of the Ishikawa line, which had been previously shown to respond to estrogen by increasing their levels of progesterone receptor and the specific activities of DNA polymerase alpha and alkaline phosphatase. Although E2 (10(-8) M) did not increase rates of proliferation during the initial logarithmic growth period of the cultures under the chosen experimental conditions (MEM with 15% charcoal-treated fetal bovine serum renewed every 2-3 days), it sustained cell proliferation after about day 10, when parallel control cultures had reached plateau cell densities. Cell proliferation in control cultures at plateau levels was resumed when the hormone was added. Growth rates of cultures containing E2 from the time of seeding and the proportion of quiescent cells, estimated by using a simple cell kinetic model, decreased steadily with time. Ornithine decarboxylase and DNA polymerase alpha activities, as well as estrogen receptor levels, also decreased with time in culture. Ishikawa cells formed colonies in soft agar; colony formation efficiencies were higher as the number of cells seeded was increased from 10,000 to 100,000 cells/6 cm dish, were not influenced by the addition of E2 to the medium (10(-9) to 10(-5) M) and were markedly reduced by difluoromethylornithine (10(-2) M), an effect that was counteracted by putrescine (25 X 10(-6) M). MH - Adenocarcinoma/METABOLISM/*PATHOLOGY ; Cell Division/DRUG EFFECTS ; Cell Line ; DNA Polymerase II/METABOLISM ; Estradiol/*PHARMACODYNAMICS ; Female ; Human ; Kinetics ; Mathematics ; Models, Biological ; Ornithine Decarboxylase/METABOLISM ; Receptors, Estrogen/METABOLISM ; Support, U.S. Gov't, P.H.S. ; Uterine Neoplasms/METABOLISM/*PATHOLOGY SO - J Steroid Biochem 1986 Nov;25(5B):781-6 3 UI - 87109508 AU - Fantini J ; Abadie B ; Tirard A ; Remy L ; Ripert JP ; el Battari A ; Marvaldi J TI - Spontaneous and induced dome formation by two clonal cell populations derived from a human adenocarcinoma cell line, HT29. AB - The replacement of glucose by galactose in the culture medium resulted in partial structural and functional enterocytic differentiation of HT29 cells. In order to characterize populations of homogeneously differentiated HT29 cells we have selected two clonal cell lines HT29-D4 and HT29-D9 with the following functional and structural characteristics when grown in a galactose-containing medium: the two clonal cell populations were permanently morphologically differentiated as shown by the presence of mature junctional complexes and a well-organized brush border (especially for HT29-D4 cells); HT29-D4 and HT29-D9 cells were able to form domes early in confluency, which indicated a functional state of differentiation; the process of differentiation was fully reversible when glucose was added to the culture medium. The induction of domes was investigated in these two cell populations and we demonstrated for the first time that proteolytic enzymes are potent inducers of dome formation. The architecture of domes either obtained spontaneously or induced by proteolytic enzymes was not maintained in the presence of ouabain (a specific inhibitor of the Na+/K+-ATPase). In conclusion, HT29-D4 and HT29-D9 cells can be maintained permanently in a differentiated state in a glucose-free medium and were able to form domes at confluency. The observation that proteolytic enzymes were able to induce dome formation can help in the comprehension of the mechanism involved in the establishment of the differentiated state. MH - Adenocarcinoma/*PATHOLOGY ; Cell Differentiation/DRUG EFFECTS ; Cell Line ; Clone Cells/ULTRASTRUCTURE ; Galactose/PHARMACODYNAMICS ; Glucose/ PHARMACODYNAMICS ; Human ; Microscopy, Electron ; Ouabain/ PHARMACODYNAMICS ; Peptide Hydrolases/PHARMACODYNAMICS ; Support, Non-U.S. Gov't SO - J Cell Sci 1986 Jul;83:235-49 4 UI - 87100997 AU - Shapiro R ; Fett JW ; Strydom DJ ; Vallee BL TI - Isolation and characterization of a human colon carcinoma-secreted enzyme with pancreatic ribonuclease-like activity. AB - A ribonuclease was isolated from serum-free supernatants of the human colon adenocarcinoma cell line HT-29. It was purified by cation-exchange and C18 reversed-phase high-performance liquid chromatography. The protein is basic, has a molecular weight of approximately 16,000, and has an amino acid composition that is significantly different from that of human pancreatic ribonuclease. The amino terminus is blocked, and the carboxyl-terminal residue is glycine. The catalytic properties of this ribonuclease resemble those of the pancreatic ribonucleases in numerous respects. Thus, it exhibits a pH optimum of approximately 6 for dinucleotide cleavage and employs a two-step mechanism in which transphosphorylation to a cyclic 2',3'-phosphate is followed by slower hydrolysis to produce a 3'-phosphate. It does not cleave NpN' substrates in which adenosine or guanosine is at the N position and prefers purines at the N' position. Like bovine ribonuclease A, the HT-29-derived ribonuclease is inactivated by reductive methylation or by treatment with iodoacetate at pH 5.5 and is strongly inhibited by the human placental ribonuclease inhibitor. However, in contrast, the tumor enzyme does not cleave CpN bonds at an appreciable rate and prefers poly(uridylic acid) as substrate 1000-fold over poly(cytidylic acid). It also hydrolyzes cytidine cyclic 2',3'-phosphate at least 100 times more slowly than uridine cyclic 2',3'-phosphate and is inhibited much less strongly by cytidine 2'-monophosphate than by uridine 2'-monophosphate. Other ribonucleases known to prefer poly(uridylic acid) were isolated both from human serum and from liver and were compared with the tumor enzyme. The physical, functional, and chromatographic properties of the serum ribonuclease are essentially identical with those of the tumor enzyme. The liver enzymes, however, differ markedly from the HT-29 ribonuclease. The potential utility of the tumor ribonuclease in the diagnosis of cancer is considered. MH - Adenocarcinoma/*ENZYMOLOGY/SECRETION ; Amino Acids/ANALYSIS ; Animal ; Cattle ; Cell Line ; Colonic Neoplasms/*ENZYMOLOGY/SECRETION ; Comparative Study ; Human ; Kinetics ; Molecular Weight ; Oligonucleotides/METABOLISM ; Pancreas/ENZYMOLOGY ; Ribonuclease, Pancreatic/*METABOLISM ; Ribonucleases/ISOLATION & PURIFICATION/ *METABOLISM ; Substrate Specificity ; Support, Non-U.S. Gov't SO - Biochemistry 1986 Nov 18;25(23):7255-64 5 UI - 87087675 AU - Bettetini D ; Garrouste F ; Remacle-Bonnet M ; Culouscou JM ; Marvaldi J ; Pommier G TI - Enhancement of production of superoxide anion by human polymorphonuclear leukocytes exposed to products of the HT-29 human colonic adenocarcinoma cell line. AB - Generation of superoxide anion (O2-) by human polymorphonuclear leukocytes (PMNs) in response to stimulation by opsonized zymosan was enhanced about 100% by prior exposure of the PMNs to human colonic adenocarcinoma cells (HT-29 cell line) or their conditioned culture medium. In addition, HT-29 cells produced substances that had an appreciable chemokinetic activity on PMNs. These tumor-secreted substances appeared to act directly on the PMNs rather than indirectly by interacting with nonadherent mononuclear cells, e.g., lymphocytes. Such a priming activity to display enhanced production of O2- was also found in conditioned medium from F344 rat FR3T3 embryonic fibroblasts but not in conditioned medium from HT-29 repolarized cells (by culture in galactose-containing medium) or from nontumorous human colonic mucosa explants. Such active substances may be important in the host-tumor relationship and, therefore, in the outcome of tumor growth. MH - Adenocarcinoma/IMMUNOLOGY/*METABOLISM ; Animal ; Cell Line ; Cell Movement ; Colonic Neoplasms/IMMUNOLOGY/*METABOLISM ; Culture Media ; Human ; Kinetics ; Lymphocytes/PHYSIOLOGY ; Neutrophils/*METABOLISM ; Oxidation-Reduction ; Phagocytosis ; Rats ; Rats, Inbred F344 ; Superoxide/*METABOLISM SO - JNCI 1986 Dec;77(6):1225-34 6 UI - 87087568 AU - Fujino N ; Kimura M ; Sakamoto K ; Shigaki N ; Yamashita J ; Akagi M TI - Tamoxifen binding sites in human mammary cancers. AB - Tamoxifen binding sites (TBS) were measured using 3H-tamoxifen, the objective being to evaluate the relationships among TBS and hormone receptors and/or clinical and pathological characteristics in malignant tissues from 60 patients with mammary cancer. TBS were detected in most (96.7 per cent) cancers in the breast tissues, and the mean content and affinity were 569 fmol/mg X protein with Kd: 1.98 nM. There was no significant correlation between TBS and the estrogen receptor and/or progesterone receptor, with respect to positivity or content. However, there was a significant correlation between TBS and histological grading, thereby indicating the differentiation and the proliferative activity in this tissue. The content of TBS was significantly higher in the group with a high grade of malignancy. The TBS content significantly increased in parallel with the degree of malignancy, as related to tubule formation, nuclear pleomorphism and mitotic activity. On the other hand, there was no significant correlation between TBS and age, tumor size, lymph node status or clinical stage. These results suggest the possibility that TBS may be associated with differentiation and cell-proliferation in breast cancer tissues. MH - Adenocarcinoma/*ANALYSIS/PATHOLOGY ; Binding Sites ; Breast Neoplasms/ *ANALYSIS/PATHOLOGY ; Carcinoma, Ductal/*ANALYSIS/PATHOLOGY ; Carcinoma, Scirrhous/*ANALYSIS/PATHOLOGY ; Cell Line ; Female ; Human ; In Vitro ; Receptors, Estrogen/*ANALYSIS ; Receptors, Progesterone/*ANALYSIS ; Tamoxifen/*ANALYSIS SO - Jpn J Surg 1986 Sep;16(5):311-7 7 UI - 87056846 AU - Yano H ; Kojiro M ; Nakashima T TI - A new human hepatocellular carcinoma cell line (KYN-1) with a transformation to adenocarcinoma. AB - A human hepatocellular carcinoma (HCC) cell line (KYN-1) has been established from a resected HCC of a 58-yr-old Japanese, male patient with HCC. Original resected HCC was moderately differentiated and proliferated in a solid pattern with vague trabecular structure in part. This cell line has been maintained for 10 mo. through 50 passages. Morphological features of KYN-1 cells demonstrated one or more large, round-to-oval nuclei with prominent nucleoli and eosinophilic polygonal-to-spindle abundant cytoplasm. In addition, some of these cells contained mucicarmin-positive materials in the cytoplasm. The cells exhibited a typical epithelial feature with pavementlike cell arrangement, and lacked contact inhibition. The doubling times of the cells grown in a serum-containing and a serum-free medium were about 31 h and 10 to 11 d, respectively. Functionally, KYN-1 cells produced albumin, alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), ferritin, beta 2-microglobulin (BMG), and alpha 1-anti-trypsin (AAT). Positive reactions for albumin, AFP, CEA, and ferritin were identified in the cells by immunohistochemical techniques. Chromosome study revealed the chromosome number in a range from 61 to 74 without mode. The tumorigenicity of KYN-1 cells was identified by the tumor formation after subcutaneous inoculation of the cells into nude mice. The developed tumor showed compact growth of the tumor cells with gland formations containing mucicarmin-positive materials. Features of adenocarcinoma were identified by electron microscopy. The tumor cells were also identified to contain albumin, AFP, CEA, ferritin, and AAT by immunohistochemical techniques. AFP, CEA, and BMG were detected in the sera of nude mice. Thus, KYN-1 cells represented the morphologic features of adenocarcinoma, retaining some characteristics of original HCC. These findings suggest that KYN-1 is a new human HCC cell line with transformation to adenocarcinoma, which will provide useful information to clarify the histogenesis of combined hepatocellular and cholangiocellular carcinoma. MH - Adenocarcinoma/*PATHOLOGY ; Alpha Fetoproteins/ANALYSIS ; Alpha 1-Antitrypsin/ANALYSIS ; Animal ; Beta 2 Microglobulin/ANALYSIS ; Carcinoembryonic Antigen/ANALYSIS ; Cell Line ; Culture Media ; Hepatoma/ ANALYSIS/*PATHOLOGY ; Human ; Liver Neoplasms/ANALYSIS/*PATHOLOGY ; Male ; Mice ; Mice, Nude ; Middle Age ; Neoplasm Transplantation ; Support, Non-U.S. Gov't ; Transplantation, Heterologous SO - In Vitro Cell Dev Biol 1986 Nov;22(11):637-46 8 UI - 87048741 AU - Yamada H ; Yoshida T ; Sakamoto H ; Terada M ; Sugimura T TI - Establishment of a human pancreatic adenocarcinoma cell line (PSN-1) with amplifications of both c-myc and activated c-Ki-ras by a point mutation. AB - A human pancreatic cancer cell line, PSN-1, was established from pancreatic adenocarcinoma tissue that had been stored for 1.5 years at -80 degrees C without any special treatment. The stored tissues were first transplanted into nude mice, and from the xenograft, the PSN-1 cell line was established. The original primary tumor and two metastatic lymph nodes were previously found to have 50-fold amplification of c-myc and also 3- to 6-fold amplification of activated c-Ki-ras with a point mutation from GGT to CGT at codon 12. PSN-1 cells are unique in that amplifications of both c-myc and activated c-Ki-ras are present in the same degree as the original tumors. These cells were also found to contain increased amounts of c-myc and c-Ki-ras transcripts. MH - Adenocarcinoma/*FAMILIAL & GENETIC/PATHOLOGY ; Animal ; Cell Line ; *Gene Amplification ; Human ; Mice ; *Mutation ; Neoplasm Transplantation ; Nucleic Acid Hybridization ; Pancreatic Neoplasms/*FAMILIAL & GENETIC/ PATHOLOGY ; *Proto-Oncogenes ; Support, Non-U.S. Gov't ; Transplantation, Heterologous SO - Biochem Biophys Res Commun 1986 Oct 15;140(1):167-73 9 UI - 87046376 AU - Kusyk CJ ; McNiel NO ; Johnson LR TI - Stimulation of growth of a colon cancer cell line by gastrin. AB - The trophic effects of the hormone gastrin-17 were examined on a human colon cancer cell line. LoVo cells were obtained from the American Type Culture Collection and grown in minimal essential medium in the presence of 10% bovine fetal serum. To demonstrate the trophic effect of gastrin, synchronization was necessary. The effect of gastrin was optimal after 26-h exposure to 0.6 mM thymidine. In the presence of serum the optimal dose of gastrin for stimulation of DNA synthesis was 7.2 X 10(-10) M. Under these conditions gastrin caused a 220% increase in [3H]thymidine incorporation. In the absence of serum the optimal dose of gastrin (3.6 X 10(-9) M) increased DNA synthesis approximately 200%. Twenty-four hours after gastrin treatment (1.8 X 10(-10) M gastrin 17) cell numbers increased 50.8% compared with control. At 48 h this increase was maintained at 44%. Maximum stimulation by gastrin occurred 7-8 h after release from synchronization and exposure to gastrin. This corresponded to the S phase of the cell cycle. Significant stimulation occurred a second time at 22-24 h, presumably during the second S phase in a still synchronous or partially synchronous cell population. These data demonstrate that physiological concentrations of gastrin-17 can stimulate the growth of a human cancer cell line and that some degree of synchronization may be necessary to demonstrate similar effects in other cell lines. Such cell lines may provide a source of rapidly growing cells in which the mechanisms of the trophic effect of gastrin can be examined. MH - Adenocarcinoma/*PATHOLOGY ; Blood ; Cell Division/DRUG EFFECTS ; Cell Line ; Colonic Neoplasms/*PATHOLOGY ; Culture Media ; DNA/BIOSYNTHESIS ; Gastrins/*PHARMACODYNAMICS ; Human ; Support, U.S. Gov't, P.H.S. SO - Am J Physiol 1986 Nov;251(5 Pt 1):G597-601 10 UI - 87045631 AU - Katsuoka Y TI - Characteristics of an established cell line (KU-2) derived from human renal cell carcinoma: I. Cloning of cells and morphological study of clones; II. Cell kinetics of KU-2 cells; III. Detection of type C virus in the culture. AB - A KU-2 cell line derived from human renal cell carcinoma was established by an indirect culture system using the nude mouse in November, 1976. These cells have been examined from different points of view including light and electron microscopic observation, and chromosomal analysis. Histopathological characteristics of the KU-2 cell line, even after being transplanted back to nude mouse, remain similar. However, the characterization of this established cell line has not been fully elucidated. In the present experiments, attempts have been made to study the cloning of KU-2 cells and morphological feature of clones, cell growth and kinetics, and detection of the type C virus in the culture. These results suggested that the KU-2 cell line was not homogeneous but composed of a heterogeneous population of cells based on morphological difference of 6 clones and discrepancy between population doubling time and generation time when calculated from the growth curve and synchronous culture of the KU-2 cells, which may be explained by the cytotoxic effect of excess thymidine. Also the type C virus was negative in the medium. MH - C-Type Viruses/*ISOLATION & PURIFICATION ; Carcinoma, Renal Cell/ MICROBIOLOGY/*PATHOLOGY ; Cell Line ; Clone Cells ; Human ; Kidney Neoplasms/MICROBIOLOGY/*PATHOLOGY ; Kinetics SO - Hinyokika Kiyo 1986 Jul;32(7):941-8 11 UI - 87030342 AU - Andrew SM ; Pimm MV ; Perkins AC ; Baldwin RW TI - Comparative imaging and biodistribution studies with an anti-CEA monoclonal antibody and its F(ab)2 and Fab fragments in mice with colon carcinoma xenografts. AB - An IgG1 mouse monoclonal antibody directed against CEA has been digested with papain to yield F(ab)2 and Fab fragments. Following radioiodination, intact antibody and fragments showed specific binding to cells of a CEA-producing tumour, although the immune reactivities of the fragments were lower than that of intact antibody. Gamma scintigraphy of nude mice bearing CEA-producing human tumour xenografts and injected with 131I-labelled fragments showed earlier and superior imaging of tumours than did 131I-intact antibody, and this was most marked with the Fab fragment. Sequential dissection analyses showed that this was due to earlier and higher tumour-to-blood ratios with fragments than with intact antibody, but in absolute terms the degree of localization of both fragment types was significantly lower than that of intact antibody. MH - Adenocarcinoma/*RADIONUCLIDE IMAGING ; Animal ; Antibodies, Monoclonal/ ANALYSIS/*DIAGNOSTIC USE/METABOLISM ; Cell Line ; Colonic Neoplasms/ *RADIONUCLIDE IMAGING ; Electrophoresis, Polyacrylamide Gel ; Human ; In Vitro ; Iodine Radioisotopes/*DIAGNOSTIC USE ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Rectal Neoplasms/*RADIONUCLIDE IMAGING ; Sarcoma, Osteogenic/*RADIONUCLIDE IMAGING ; Support, Non-U.S. Gov't ; Time Factors ; Tissue Distribution SO - Eur J Nucl Med 1986;12(4):168-75 12 UI - 87011809 AU - deKernion JB ; Lovrekovich L ; Chopin D ; Studer UE TI - Antibodies to cultured tumor cells detected in sera of renal cell carcinoma patients by a quantitative avidin-biotin method. AB - Antibodies reacting with the tumor cell line RC-Pa were measured by a quantitative avidin-biotin complex method. Sera of renal cell carcinoma patients, patients with other types of cancer and healthy donors were analyzed. Of 71 sera from renal cell carcinoma patients 67 (94 per cent) were classified as showing renal cell carcinoma, while 32 of 36 sera (89 per cent) from healthy subjects were classified as showing no renal cell carcinoma. Four of 21 serum specimens (19 per cent) from individuals with other than renal cancer were misclassified. Furthermore, sera from renal carcinoma patients immunized with a mixture of autologous tumor cells and Corynebacterium parvum showed a marked increase in reactivity compared to those from patients receiving progesterone. The results indicate that this assay might become useful to detect or monitor renal cell carcinoma. MH - Antibodies, Neoplasm/*ANALYSIS ; Avidin/*DIAGNOSTIC USE ; Biotin/ *DIAGNOSTIC USE ; Carcinoma, Renal Cell/*IMMUNOLOGY ; Cell Line ; Female ; Human ; Kidney Neoplasms/*IMMUNOLOGY ; Male ; Middle Age ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. ; Tumor Stem Cell Assay SO - J Urol 1986 Oct;136(4):795-8 13 UI - 87008731 AU - Saksela K ; Bergh J ; Nilsson K TI - Amplification of the N-myc oncogene in an adenocarcinoma of the lung. AB - c-myc oncogene is the most extensively studied member of the myc gene family, which now consists of three characterized members, namely the c-myc, N-myc, and L-myc genes. Deregulation owing to amplification and/or rearrangements of the c-myc gene have been described in a variety of human malignancies. Several neuroblastomas have amplifications of the N-myc genes. The c-myc, N-myc, or L-myc oncogenes are also found amplified in different cell lines from small cell carcinomas of the lung. In this study, we have examined the c-myc, N-myc, and c-erbB oncogenes in 34 clinical and autopsy tumor specimens representing various histopathological types of human lung cancer, including nine small cell lung cancers. A 30-fold amplification of the N-myc gene was found in a tumor histopathologically and histochemically verified as a typical adenocarcinoma. No amplifications of the c-myc or c-erbB oncogenes were seen in any of the tumors. In the DNA of one small cell carcinoma, an extra c-myc and N-myc cross-hybridizing restriction fragment was observed, possibly owing to an amplification of a yet uncharacterized myc-related gene. MH - Adenocarcinoma/*FAMILIAL & GENETIC ; Carcinoma, Oat Cell/FAMILIAL & GENETIC ; Cell Line ; DNA, Neoplasm/GENETICS ; *Gene Amplification ; Human ; Lung Neoplasms/*FAMILIAL & GENETIC ; *Oncogenes ; Support, Non-U.S. Gov't SO - J Cell Biochem 1986;31(4):297-304 14 UI - 87007184 AU - Oosterwijk E ; Ruiter DJ ; Hoedemaeker PJ ; Pauwels EK ; Jonas U ; Zwartendijk J ; Warnaar SO TI - Monoclonal antibody G 250 recognizes a determinant present in renal-cell carcinoma and absent from normal kidney. AB - This report describes a monoclonal antibody designated G 250, subclass IgGl, that recognizes an antigen preferentially expressed on cell membranes of renal-cell carcinoma cells (RCC) and not expressed in normal proximal tubular epithelium. G 250 antibody reacted with 46 of 47 primary RCC, with 7 of 8 RCC metastases and with a few other malignant tumors. The staining pattern of G 250 differs from that of other RCC-related antibodies described. Preliminary experiments show that this antibody can be used to visualize RCC xenografts in nude mice by immunoscintigraphy. MH - Animal ; *Antibodies, Monoclonal ; Antigenic Determinants/*IMMUNOLOGY ; Carcinoma, Renal Cell/*IMMUNOLOGY ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Epithelium/IMMUNOLOGY ; Human ; Kidney Tubules, Proximal/CYTOLOGY/IMMUNOLOGY ; Kidney/*IMMUNOLOGY ; Mice ; Mice, Nude ; Radionuclide Imaging ; Tissue Distribution ; Transplantation, Heterologous SO - Int J Cancer 1986 Oct 15;38(4):489-94 15 UI - 87002865 AU - Scher HI ; Sternberg C ; Heston WD ; Watson RC ; Niedzwiecki D ; Smart T ; Hollander P ; Yagoda A TI - Etoposide in prostatic cancer: experimental studies and phase II trial in patients with bidimensionally measurable disease. AB - Etoposide, a semisynthetic derivative of podophyllotoxin, was evaluated concurrently in vitro against a human derived hormone-resistant cell line, PC-3, and in vivo in bidimensionally measurable hormone-resistant human prostatic cancer. In vitro, a dose-response relationship was observed, with 74% inhibition at 10 micrograms/ml (1 h incubation) and greater than 99% inhibition at 90 micrograms/ml, both in the range of clinically achievable concentrations. In vivo, 1 PR (5%, 95% confidence limits 0-12%) of 18+ months was observed in 20 adequately treated patients. The results confirm the limited role of etoposide in hormone-refractory disease and the need for new model systems for evaluation of potential chemotherapeutic compounds in this disease. MH - Adenocarcinoma/*DRUG THERAPY ; Cell Line ; Drug Evaluation ; Drug Resistance ; Etoposide/*THERAPEUTIC USE ; Hormones/THERAPEUTIC USE ; Human ; Male ; Prostatic Neoplasms/*DRUG THERAPY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Tumor Stem Cell Assay SO - Cancer Chemother Pharmacol 1986;18(1):24-6 16 UI - 87002098 AU - Friedman EL ; Hayes DF ; Kufe DW TI - Reactivity of monoclonal antibody DF3 with a high molecular weight antigen expressed in human ovarian carcinomas. AB - We have previously described the monoclonal antibody (MAb) DF3, prepared against a human breast carcinoma. MAb DF3 reacts with a high molecular weight glycoprotein detectable in human breast carcinomas and in human milk. Previous studies have demonstrated that DF3 antigen levels are elevated in the plasma of patients with breast and ovarian cancer. The present study has further examined the reactivity of MAb DF3 with human ovarian carcinomas. Immunoperoxidase staining demonstrated reactivity of MAb DF3 with 95% of benign, borderline, and malignant tumors (serous, mucinous, and endometrioid) of the ovary. Furthermore, malignant tumors contained cytoplasmic DF3 antigen while benign tumors expressed the antigen only on apical surfaces. Western blot analyses demonstrated that the MAb DF3 reactive ovarian antigen (DF3-O) was a glycoprotein with a heterogenous molecular weight ranging between 300,000 and 450,000. This antigen was detectable by immunofluorescence on the cell surface of five of six cultured human ovarian carcinoma cell lines. The extent of cell surface reactivity with MAb DF3 was equivalent to or greater than that obtained with MAb OC125, an antibody generated against coelomic epithelium and developmental amnion. Furthermore, uptake of 125I-labeled MAb DF3 by human ovarian carcinoma xenografts in athymic mice was 5.4- and 6.2-fold higher than the respective uptake noted in liver and control tumor (P = 0.031). These findings suggest that DF3-O antigen is similar if not identical to the antigen detected in human breast carcinomas by MAb DF3. Thus, MAb DF3 may be a useful reagent in immunodiagnostic evaluation of patients with ovarian cancer. MH - Adenocarcinoma/*IMMUNOLOGY ; Animal ; Antibodies, Monoclonal/*IMMUNOLOGY ; Antigens, Neoplasm/*ANALYSIS ; Antigens, Surface/ANALYSIS ; Breast Neoplasms/ANALYSIS ; Cell Line ; Female ; Human ; Immunoenzyme Technics ; Mice ; Mice, Inbred BALB C ; Molecular Weight ; Ovarian Neoplasms/ *IMMUNOLOGY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Cancer Res 1986 Oct;46(10):5189-94 17 UI - 86317309 AU - Katsuoka Y ; Hoshino H ; Shiramizu M ; Sakabe K ; Seiki K TI - Autoradiographic and cytochemical localization of androgen in human prostatic cancer cell lines. AB - For basic studies of receptor dynamics in androgen-responsive tissues and cells, the autoradiographic and cytochemical procedures were applied to cultured tumor cells (DU-145 and PC-3). Uptake and retention of 3H-R1881, a potent synthetic androgen, were observed in DU-145 cells. The radioactive labelling was intense, and solely confined to the nuclei of DU-145 cells. Radioactivity over PC-3 cells was minimal. For assessing binding specificity, DU-145 cells were incubated with 3H-R1881 in the presence or absence of either unlabelled R1881, testosterone, progesterone, estradiol-17 beta, or corticosterone. The displacement of 3H-R1881 with R1881 and testosterone was significant, while no displacement was observed with other steroids. Nuclear localization of cytochemical staining of the dihydrotestosterone-peroxidase conjugate was evident in DU-145 cells. Our results indicate that androgen receptor may reside primarily in target cell nuclei of androgen-responsive tissues and tumors. MH - Adenocarcinoma/*METABOLISM ; Autoradiography ; Binding, Competitive ; Cell Line ; Estrenes/DIAGNOSTIC USE/METABOLISM ; Histocytochemistry ; Human ; Male ; Prostatic Neoplasms/*METABOLISM ; Receptors, Androgen/ *METABOLISM SO - Urology 1986 Sep;28(3):228-31 18 UI - 86308824 AU - Chin JK ; Rong GH ; Scharff JE ; Sindelar WF TI - Gastrointestinal carcinoma-associated antigen defined by a murine monoclonal antibody. AB - A murine monoclonal antibody, CHIP, has been prepared against a human pancreatic carcinoma cell line, SHAW. With the use of the avidin-biotin immunoperoxidase technique, the CHIP antibody detected an antigen found in 11 of 20 fixed tissue sections of tumors obtained from patients with pancreatic carcinoma. The antibody also detected the antigen in 25 of 26 colon carcinoma specimens, 4 of 6 gastric carcinoma specimens, and 1 of 2 esophageal adenocarcinoma specimens. The antigen was also found in normal proximal jejunum and colon and in small amounts in pancreatic islets and parathyroid. There was no reactivity with normal pancreatic ductal or acinar cells or with mesenchymal tissues. MH - Adenocarcinoma/*IMMUNOLOGY ; Animal ; Antibodies, Monoclonal/*IMMUNOLOGY ; Antigens, Neoplasm/*ANALYSIS ; Carcinoembryonic Antigen/IMMUNOLOGY ; Cell Line ; Gastrointestinal Neoplasms/*IMMUNOLOGY ; Human ; Immunoenzyme Technics ; Mice ; Mice, Inbred BALB C ; Pancreatic Neoplasms/*IMMUNOLOGY SO - JNCI 1986 Sep;77(3):599-604 19 UI - 86291136 AU - Yanagawa T ; Hayashi Y ; Nagamine S ; Yoshida H ; Yura Y ; Sato M TI - Generation of cells with phenotypes of both intercalated duct-type and myoepithelial cells in human parotid gland adenocarcinoma clonal cells grown in athymic nude mice. AB - A neoplastic epithelial cell line was established from nude mice tumors grown after transplantation of surgical specimens from a human parotid gland adenocarcinoma. This cell line, which had ultrastructure similarities to salivary intercalated duct cells, was found by immunohistochemical techniques to contain amylase, but myosin was not detected. Ultrastructurally, cells of an intermediate type between intercalated ductal and myoepithelial cells were found in the transplanted tumors. Moreover, the expression of myosin in addition to the presence of amylase was detected in the tumors. These findings indicate that some transplanted tumor cells appear to be differentiating towards myoepithelial cells. MH - Adenocarcinoma/ENZYMOLOGY/*PATHOLOGY ; Amylases/ANALYSIS ; Animal ; Cell Line ; Human ; Immune Sera ; Immunoenzyme Technics ; Mice ; Mice, Nude ; Myosin/ANALYSIS ; Neoplasm Transplantation ; Parotid Neoplasms/ENZYMOLOGY/ *PATHOLOGY ; Phenotype ; Support, Non-U.S. Gov't ; Transplantation, Heterologous SO - Virchows Arch [B] 1986;51(3):187-95 20 UI - 86290368 AU - Kotanagi H ; Takahashi T ; Masuko T ; Hashimoto Y ; Koyama K TI - A monoclonal antibody against human colon cancers. AB - A monoclonal antibody was prepared by hybridizing mouse myeloma cells with spleen cells from the mouse which was immunized with human colon cancer transplanted in nude mice. The reactivity of the monoclonal antibody, named A7, was tested by immunoperoxidase method. A7 reacted strongly with human adenocarcinoma cell lines and carcinoembryonic antigen (CEA). In surgical specimens, A7 reacted with 10 cancer tissues and 2 normal colon mucosa from 19 colorectal cancer patients. A7 did not react with other cancers. It was thought that A7 reacted with colon- or colon cancer-specific CEA. The reactivity of A7 with colorectal cancers was markedly reduced by preoperative irradiation. MH - Adenocarcinoma/*IMMUNOLOGY ; Adult ; Aged ; Animal ; Antibodies, Monoclonal/*IMMUNOLOGY ; Antigens, Neoplasm/*IMMUNOLOGY ; Carcinoembryonic Antigen/IMMUNOLOGY ; Cell Line ; Colonic Neoplasms/ *IMMUNOLOGY ; Female ; Human ; Immunoenzyme Technics ; Male ; Mice ; Mice, Nude ; Middle Age ; Rectal Neoplasms/*IMMUNOLOGY ; Support, Non-U.S. Gov't SO - Tohoku J Exp Med 1986 Apr;148(4):353-60 21 UI - 86281954 AU - Ripoll EA ; Rama BN ; Webber MM TI - Vitamin E enhances the chemotherapeutic effects of adriamycin on human prostatic carcinoma cells in vitro. AB - Vitamin E (tocopherol) enhances the growth inhibitory effects of adriamycin (ADR) on a variety of cancer cells in vitro. The role of vitamin E (d-alpha-tocopheryl) acid succinate in adjuvant chemotherapy with ADR was assessed in DU-145 human prostatic carcinoma cells in culture. Adriamycin produced a dose-dependent growth inhibition of DU-145 cells. The ID50 of DU-145 cells on the criteria: of clonal assay was 13 ng./ml. and of cell count assay was 14 ng./ml. Vitamin E succinate also inhibited the growth of DU-145 human prostatic carcinoma cells in a dose-dependent manner. 4.4 micrograms./ml. and 5.4 micrograms./ml. vitamin E succinate in the culture medium produced inhibition of growth to 50 per cent of control (ID50) in the clonal and the cell count assays respectively. When adriamycin and vitamin E succinate were used in combination, both additive and synergistic effects were observed, depending on the concentration of vitamin E succinate used. Doses of vitamin E succinate greater than its ID50 had a synergistic effect while doses smaller than its ID50 had an additive effect. In either case, the presence of vitamin E succinate caused an enhancement of tumor cell cytotoxicity of adriamycin while decreasing its ID50. Equivalent concentrations of sodium succinate and ethanol used to dissolve vitamin E succinate did not have any effect on DU-145 cells. Thus, it is concluded that the effect of vitamin E succinate is due to vitamin E and not due to succinate or ethanol. These results suggest that vitamin E may have a role in the treatment of human prostatic cancer as an adjuvant agent to adriamycin. MH - Adenocarcinoma/*PATHOLOGY ; Cell Line ; *Colony-Forming Units Assay ; Dose-Response Relationship, Drug ; Doxorubicin/*PHARMACODYNAMICS ; Drug Synergism ; Human ; In Vitro ; Male ; Prostatic Neoplasms/*PATHOLOGY ; Support, U.S. Gov't, P.H.S. ; *Tumor Stem Cell Assay ; Vitamin E/ANALOGS & DERIVATIVES/*PHARMACODYNAMICS SO - J Urol 1986 Aug;136(2):529-31 22 UI - 86277145 AU - Jenkins VK ; Barranco SC ; Townsend CM Jr ; Perry RR ; Ives KL TI - Differential response to gamma radiation of human stomach cancer cells in vitro. AB - In vitro effects of radiation were studied in two permanent cell lines (AGS and SII) from two patients with adenocarcinoma of the stomach and three permanent sublines from each cell line. Radiation survival parameters for AGS and SII parent cell lines and sublines were determined after in vitro irradiation of their cells with 0.5 to 10 Gy of 60Co gamma rays. The AGS and SII cell lines had different growth properties, DNA contents and radiation survival curves. Surviving fractions of SII parent cells (76 chromosomes) after 2.0 and 10 Gy were 1.22 and 17.8 times greater, respectively, than values for AGS parent cells (47 chromosomes). Sensitivities (D0) were 1.08 and 1.45 Gy for AGS and SII parent lines, respectively. The D0 values for AGS parent cells and sublines were similar (1.01 to 1.08 Gy), but SII parent cells and sublines had D0 values of 1.45, 1.36, 1.37 and 1.12 Gy (for SII-A). Also, the SII parent cells had survival fractions after 2.0 and 10 Gy that were 1.3 and 11.3 times greater, respectively, than values for the SII-A cells. These data show differences in radiation responses among stomach cancer cell lines and sublines that may relate to DNA content, but there was no consistent correlation between radiation response and a particular cell characteristic. MH - Adenocarcinoma/PATHOLOGY/*RADIOTHERAPY ; Cell Cycle/RADIATION EFFECTS ; Cell Line ; Cell Survival/RADIATION EFFECTS ; Cells, Cultured ; Chromosomes, Human/ULTRASTRUCTURE ; DNA, Neoplasm/ANALYSIS/RADIATION EFFECTS ; Gamma Rays/THERAPEUTIC USE ; Human ; Stomach Neoplasms/ PATHOLOGY/*RADIOTHERAPY ; Support, U.S. Gov't, P.H.S. ; Time Factors SO - Int J Radiat Biol 1986 Aug;50(2):269-78 23 UI - 86276192 AU - Ishiwata I ; Ishiwata C ; Soma M ; Ishikawa H TI - Establishment and characterization of two human ovarian endometrioid carcinoma cell lines (with or without squamous cell component). AB - The cell lines designated HMOA and HNOA were established from human ovarian adenoacanthoma and from a mouse graft of human ovarian endometrioid adenocarcinoma, respectively. These cell lines grew well without interruption for over 20 months. The cultured cells of both HMOA and HNOA lines were spindle, polygonal, and columnar, and showed a jigsaw puzzle-like arrangement and a piling-up tendency devoid of contact inhibition. When the HMOA cells were maintained at the confluent stage, the cells formed cysts and/or squamous metaplasia. The chromosome number of both cell lines varied widely and showed aneuploidy, while the modal chromosome number was stable at the diploid range. Both of these cell lines, HMOA and HNOA, were transplanted into the subcutis of BALB/c nude mice and produced well-differentiated adenoacanthoma and poorly differentiated endometrioid adenocarcinoma, respectively. HMOA cells were characterized as producing large amounts of CA125 (ovarian carcinoma marker), in vitro, in the cyst-forming phase. The HNOA cells, however, did not produce CA125. MH - Adenocarcinoma/*PATHOLOGY ; Aged ; Animal ; Cell Line ; Female ; Human ; Karyotyping ; Mice ; Mice, Inbred BALB C ; Middle Age ; Neoplasm Transplantation ; Ovarian Neoplasms/*PATHOLOGY ; Support, Non-U.S. Gov't SO - Gynecol Oncol 1986 Sep;25(1):95-107 24 UI - 86274888 AU - Houghton JA ; Torrance PM ; Radparvar S ; Williams LG ; Houghton PJ TI - Binding of 5-fluorodeoxyuridylate to thymidylate synthase in human colon adenocarcinoma xenografts. AB - The formation and stability of the covalent ternary complex formed between thymidylate synthase (E.C. 2.1.1.45), 5-fluoro 2'-deoxyuridylate (FdUMP) and 5,10-methylenetetrahydrofolate (CH2-H4PteGlu) has been examined in cytosols derived from xenografts of human colon adenocarcinomas. The rate of association (ka) for FdUMP was low being between 3.4 +/- 0.9 and 10.2 +/- 2.6 X 10(6) M-1 min-1, with the lowest ka value being determined in cytosols from a tumor (HxELC2) which has demonstrated some sensitivity to 5-fluoropyrimidines. Relative to reported ka values for human leukemic cells, the rate of association of FdUMP was 20- to 59-fold lower. This difference is not a consequence of FdUMP catabolism, or metabolism of CH2-H4PteGlu. In cytosols the apparent Km values for dUMP (3.6-4.2 microM) and and [6RS]- CH2-H4PteGlu (25-26.7 microM) were similar to reported values for human enzyme. Data derived from cytosols were similar to those derived using affinity purified enzyme from HxVRC5 colon adenocarcinoma xenografts. The net dissociation of [6-3H] FdUMP from the covalent ternary complex was 31-33 min in the absence of added CH2-H4PteGlu, and the rate of dissociation was dependent upon the concentration of cofactor. The concentration of [6RS]-CH2-H4PteGlu required to stabilize ternary complex derived from HxELC2 cytosols was slightly lower than that required for the same degree of stabilization of complex formed in cytosols from resistant tumors (HxGC3,HxVRC5). Addition of 5-CHO-H4PteGlu, 5-CH3-H4PteGlu, H2PteGlu, and PteGlu did not stabilize the covalent complex, but H4PteGlu substituted for CH2-H4PteGlu. MH - Adenocarcinoma/*METABOLISM ; Animal ; Cell Line ; Colonic Neoplasms/ *METABOLISM ; Deoxyuracil Nucleotides/*METABOLISM ; Fluorodeoxyuridylate/ *METABOLISM ; Human ; Kinetics ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Thymidylate Synthetase/*METABOLISM ; Transplantation, Heterologous SO - Eur J Cancer Clin Oncol 1986 Apr;22(4):505-10 25 UI - 86271768 AU - Sobol RE ; Peters RE ; Astarita RW ; Hofeditz C ; Masui H ; Burton D ; Handley HH ; Glassy MC ; Fairshter R ; Carlo DJ ; et al TI - A novel monoclonal antibody-defined antigen which distinguishes human non-small cell from small cell lung carcinomas. AB - Spleen cells from BALB/c mice hyperimmunized with the human epidermoid lung carcinoma cell line T222 were fused with NS-1 mouse myeloma cells to produce monoclonal antibodies to human lung cancer antigens. Hybridoma culture supernatants were tested by an enzyme-linked immunosorbent assay for reactivity against a panel of human lung tumor cell lines. Supernatant from hybridoma EA1 (immunoglobulin G1) displayed strong reactivity with four of four non-small cell lung carcinomas but did not react with three of three small cell lung carcinoma (SCLC) cell lines. This hybridoma was cloned by limiting dilution and utilized to generate ascites antibody for subsequent immunohistochemical and antigen characterization studies. Evaluation of fresh frozen tumor tissue sections by immunoperoxidase staining methods revealed EA1 reactivity with the vast majority of non-SCLCs tested (21 of 21 epidermoid, 17 of 18 adenocarcinomas, four of four large cell, two of two bronchioloalveolar) and no reactivity with nine of nine small cell lung carcinomas. EA1 also stained bronchial epithelium and other benign and malignant epithelial tissues. The EA1 antigen was determined to have a molecular weight of 75,000 by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of human non-SCLC tumor extracts. These data imply that EA1 recognizes a novel antigen expressed by non-SCLCs and other epithelial tissues. The absence of EA1 reactivity with SCLCs suggests that this monoclonal antibody may find future application in distinguishing non-SCLC from SCLC and prove useful in furthering our understanding of the histogenesis of lung carcinomas. MH - Adenocarcinoma/*IMMUNOLOGY ; Antibodies, Monoclonal/*IMMUNOLOGY ; Antigens, Neoplasm/*IMMUNOLOGY ; Antigens, Surface/IMMUNOLOGY ; Carcinoma, Oat Cell/*IMMUNOLOGY ; Carcinoma, Squamous Cell/*IMMUNOLOGY ; Cell Line ; Cell Membrane/IMMUNOLOGY ; Epithelium/IMMUNOLOGY ; Human ; Lung/IMMUNOLOGY ; Lung Neoplasms/*IMMUNOLOGY ; Molecular Weight ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. SO - Cancer Res 1986 Sep;46(9):4746-50 26 UI - 86271760 AU - O'Hara BM ; Oskarsson M ; Tainsky MA ; Blair DG TI - Mechanism of activation of human ras genes cloned from a gastric adenocarcinoma and a pancreatic carcinoma cell line. AB - We have analyzed the mechanism of activation of two human ras oncogenes. We have also identified a rasN gene from a human gastric adenocarcinoma which efficiently induced both morphological transformation and tumorigenicity of NIH3T3 cells in a transfection assay. The rasN gene in tumor tissue DNA did not appear to be rearranged or amplified. A molecular clone, which contained an EcoRI fragment spanning the first and second rasN exons, was molecularly cloned directly from the human tumor DNA. Chimeric constructions and DNA sequencing defined the mechanism of activation of the gene as a mutation in the 61st amino acid codon substituting arginine for glutamine. Normal DNA isolated from Epstein-Barr virus immortalized lymphocytes derived from the same patient did not induce morphological transformation or tumorigenicity in NIH3T3 cells. A cloned cell line isolated from the human pancreatic carcinoma cell line Panc1 had previously been shown to contain an activated rasK-2. Sequence analysis of the cloned transfected gene reveals a G to A change within codon 12, which is presumably responsible for its biological activity. This represents the first identification of a codon 12 aspartic acid substitution of a c-rask oncogene from a human tumor-derived cell line. MH - Adenocarcinoma/*FAMILIAL & GENETIC ; Amino Acid Sequence ; Carcinoma/ *FAMILIAL & GENETIC ; Cell Line ; Cloning, Molecular ; DNA, Neoplasm/ *GENETICS ; Gene Expression Regulation ; Human ; *Oncogenes ; Pancreatic Neoplasms/*FAMILIAL & GENETIC ; Stomach Neoplasms/*FAMILIAL & GENETIC ; Support, U.S. Gov't, P.H.S. SO - Cancer Res 1986 Sep;46(9):4695-700 27 UI - 86271716 AU - Hanai N ; Shitara K ; Yoshida H TI - Generation of monoclonal antibodies against human lung squamous cell carcinoma and adenocarcinoma using mice rendered tolerant to normal human lung. AB - Four murine monoclonal antibodies against human lung carcinoma were generated using a novel immunization procedure. BALB/c mice were rendered neonatally tolerant to normal human lung tissues and subsequently immunized with human lung tumor tissues. The lower level of antibody-reactivity to the tolerogen was seen in the sera of mice rendered neonatally tolerant as compared with the level of reactivity in the sera of nontolerant mice. The mice which maintained a sufficiently tolerant state were selected for hybridoma production. Two monoclonal antibodies, KM-32 and KM-34, were developed from mice immunized with lung squamous cell carcinoma tissues and two other monoclonal antibodies, KM-52 and KM-93, were developed from mice immunized with lung adenocarcinoma tissues. Distribution of antigens detected by the monoclonal antibodies were investigated by enzyme-linked immunosorbent assay using membrane fractions prepared from a number of tumorous and normal tissues and various human normal and tumor cell lines. KM-32 recognized a carbohydrate antigen expressed predominantly on lung squamous cell carcinoma cells and its carbohydrate structure appeared to be associated with blood group A antigen. KM-93 recognized a sialylated carbohydrate epitope on the antigen expressed on lung adeno-carcinoma cells and a few other tumor cells. KM-34 and KM-52 detected protein or glycoprotein antigens and they showed predominant reactivity to lung squamous cell carcinoma and lung adenocarcinoma, respectively. KM-32 and KM-34 antibodies showed complement-dependent cytotoxicity against a lung tumor cell line. These results suggest that the tolerance technique is useful for efficient screening of murine monoclonal antibodies specific to human tumors. The efficacy of these four monoclonal antibodies in diagnosis and therapy of lung cancer will be the subject of a sequential study. MH - Adenocarcinoma/*IMMUNOLOGY ; Animal ; Antibodies, Monoclonal/*IMMUNOLOGY ; Antibody Specificity ; Binding, Competitive ; Carcinoma, Squamous Cell/ *IMMUNOLOGY ; Cell Line ; Cell Membrane/IMMUNOLOGY ; Cytotoxicity Tests, Immunologic ; Enzyme-Linked Immunosorbent Assay ; Hemagglutination Tests ; Human ; Immune Tolerance ; Lung/IMMUNOLOGY ; Lung Neoplasms/*IMMUNOLOGY ; Mice SO - Cancer Res 1986 Sep;46(9):4438-43 28 UI - 86271636 AU - Baumal R ; Law J ; Buick RN ; Kahn H ; Yeger H ; Sheldon K ; Colgan T ; Marks A TI - Monoclonal antibodies to an epithelial ovarian adenocarcinoma: distinctive reactivity with xenografts of the original tumor and a cultured cell line. AB - Four monoclonal antibodies (mAb) (8C, 10B, M2A, and M2D) were produced against the human epithelial ovarian adenocarcinoma cell line, HEY. The affinity constants of binding of the mAb to cultured HEY cells were 8 X 10(8) M-1 (M2D) and 10(9) M-1 (8C and 10B). mAb 8C reacted with a major glycoprotein of Mr 90,000 on the surface of HEY cells. The four mAb differed from previously reported mAb to epithelial ovarian adenocarcinomas on the basis of their reactivity with cultured ovarian adenocarcinoma cell lines using a cell-binding radioimmunoassay, and their staining of cryostat sections of various human normal and tumor tissues using an immunoperoxidase reaction. All four mAb reacted with s.c. tumors derived by injecting cultured HEY cells into thymectomized CBA/CJ mice. However, only two of the four mAb (8C and 10B) also reacted with s.c. tumors of the original HEY xenograft from which the cultured cell line was derived. In addition, mAb 8C and 10B reacted by immunoperoxidase staining with 2 and 4 different cases, respectively, of 11 epithelial ovarian adenocarcinomas examined. Cultured HEY cells were adapted to grow i.p. in BALB/c-nu/nu mice and the i.p. tumors retained their reactivity with the monoclonal antibodies. These tumor-bearing mice offer a useful model system for studying the potential of mAb, especially 8C and 10B, for the diagnosis and treatment of patients with peritoneal extension of epithelial ovarian adenocarcinomas. MH - Adenocarcinoma/*IMMUNOLOGY/THERAPY ; Animal ; Antibodies, Monoclonal/ *IMMUNOLOGY/THERAPEUTIC USE ; Cell Line ; Female ; Human ; Kidney/ IMMUNOLOGY ; Mice ; Mice, Inbred BALB C ; Mice, Inbred CBA ; Neoplasm Transplantation ; Ovarian Neoplasms/*IMMUNOLOGY/THERAPY ; Rabbits ; Support, Non-U.S. Gov't ; Transplantation, Heterologous SO - Cancer Res 1986 Aug;46(8):3994-4000 29 UI - 86260794 AU - Streffer C ; van Beuningen D ; Gross E ; Schabronath J ; Eigler FW ; Rebmann A TI - Predictive assays for the therapy of rectum carcinoma. AB - Cytofluorometric DNA measurements showed that about 55% of rectum carcinoma (129 patients) had tumours with an abnormal DNA content (DNA aneuploidy). For patients with such a tumour the prognosis was worse than for patients with DNA diploid tumours. From the DNA histograms the number of S-phase cells was calculated. In tumours with the stage pT3, which disseminated to lymph nodes or metastasized, a higher number of S-phase cells was found than in tumours with the staging pT3N0M0. In all untreated tumours cells with micronuclei were found. This demonstrated cell loss. In most tumours this effect was considerable. The ratio:number of S-phase cells/number of cells with micronuclei may allow a rough estimate for cell turnover. In patients with a bad prognosis and in those patients who had a local recurrence after resection of the tumour this ratio was high. In 34 patients the parameters were measured before and after preoperative radiotherapy. In some tumours a rapid increase of S-phase cells occurred after irradiation, this effect might express repopulation. In these patients a local recurrence was frequently found. From the data obtained so far a prediction for local recurrences might be possible from the determination of nuclear protein bound SH-groups. The determination of micronuclei indicated that it can be used as a measure for radiation response in tumours. All parameters show a high variability between individual tumours. A further study is useful whether the measured parameters are suitable as predictors. MH - Adenocarcinoma/PATHOLOGY/*RADIOTHERAPY ; Aneuploidy ; Biopsy ; Cell Line ; Cell Nucleus/*RADIATION EFFECTS ; Combined Modality Therapy ; *Cytophotometry ; DNA, Neoplasm/*METABOLISM ; Human ; Interphase/ RADIATION EFFECTS ; Neoplasm Recurrence, Local/ETIOLOGY ; Neoplasm Staging ; Prognosis ; Rectal Neoplasms/PATHOLOGY/*RADIOTHERAPY ; Support, Non-U.S. Gov't SO - Radiother Oncol 1986 Apr;5(4):303-10 30 UI - 86255593 AU - Lundy J ; Greiner JW ; Colcher D TI - Development of a metastatic human colon cancer xenograft model in the nude mouse. AB - The objective of our experimental protocols was to develop a metastatic model for a human colon carcinoma xenograft in congenitally athymic nude mice. This model would be useful in evaluating the efficacy of radiolabeled monoclonal antibodies for detection and treatment of regional and distant metastases. The LS-174T human colon carcinoma line was used to establish primary subcutaneous tumors in nude mice. Mice were killed at varying time intervals to establish the incidence of spontaneous metastases. Only lung micrometastases were observed during the 2-month observation period. To increase the metastatic rate, the site of primary implantation was varied and/or surgical manipulations were performed. Excision of small primary tumors resulted in a low incidence of local recurrence and no distant metastases. However, with excision of large primary tumors, a high local recurrence rate was noted and over 30% of mice had gross metastases. Mice bearing hind footpad tumors underwent excision when tumors were at least 1 cm in size. There were no local recurrences, but by 8 weeks over 40% had large pulmonary metastases. The LS-174T tumor was also established as a primary implant in the spleen from which 10 to 15% of the mice developed liver or lung metastases. The LS-174T tumor can metastasize in the nude mice and the latter two models may prove very useful in imaging and therapy studies. MH - Adenocarcinoma/PATHOLOGY/*SECONDARY ; Animal ; Antibodies, Monoclonal/ DIAGNOSTIC USE ; Cell Line ; Colonic Neoplasms/*PATHOLOGY ; Human ; Liver Neoplasms, Experimental/PATHOLOGY ; Lung/PATHOLOGY ; Lung Neoplasms/ PATHOLOGY/SECONDARY ; Male ; Mice ; *Mice, Nude ; Muscles/PATHOLOGY ; Neoplasm Metastasis ; Neoplasm Recurrence, Local/PATHOLOGY ; Neoplasm Transplantation ; Spleen/PATHOLOGY ; Splenic Neoplasms/PATHOLOGY/ SECONDARY ; Transplantation, Heterologous SO - J Surg Oncol 1986 Apr;31(4):260-7 31 UI - 86251176 AU - Gravanis A ; Gurpide E TI - Effects of estradiol on deoxyribonucleic acid polymerase alpha activity in the Ishikawa human endometrial adenocarcinoma cell line. AB - The effects of estradiol on DNA polymerase alpha activity were investigated in an estrogen-responsive human endometrial cancer cell line (Ishikawa) derived from a well differentiated endometrial adenocarcinoma. These cells are known to respond to estradiol by increasing progesterone receptor levels, alkaline phosphatase activity, and cell density. Four- to 5-fold increases in DNA polymerase alpha activity occurred when estradiol was added to cultures of Ishikawa cells in medium containing charcoal-treated fetal bovine serum. Maximal stimulation was achieved at 18 h during incubations with 10(-8) M estradiol, but significant effects also were found with 10(-9) and 10(-6) M. These effects were almost completely counteracted by a 100-fold excess of 4-hydroxytamoxifen. At 10(-6) M, the antiestrogen had no influence on the basal levels of DNA polymerase alpha. Medroxyprogesterone acetate (10(-6) M) was ineffective as either an enhancer of enzymatic activity or an antiestrogen when tested in combination with 10(-8) M estradiol, even in the presence of appreciable levels of specific progesterone binders. The responsiveness of the Ishikawa cells to estrogen contrasts with the lack of effects of estradiol on DNA polymerase alpha activity in another human endometrial adenocarcinoma cell line (HEC-50). MH - Adenocarcinoma/*ENZYMOLOGY ; Cell Line ; DNA Polymerase II/ANTAGONISTS & INHIBITORS/*METABOLISM ; Enzyme Activation/DRUG EFFECTS ; Estradiol/ *PHARMACODYNAMICS ; Female ; Human ; Medroxyprogesterone/ANALOGS & DERIVATIVES/PHARMACODYNAMICS ; Support, U.S. Gov't, P.H.S. ; Tamoxifen/ ANALOGS & DERIVATIVES/PHARMACODYNAMICS ; Uterine Neoplasms/*ENZYMOLOGY SO - J Clin Endocrinol Metab 1986 Aug;63(2):356-9 32 UI - 86251077 AU - el-Battari A ; Muller JM ; Fantini J ; Bellot F ; Tirard A ; Ducret F ; Marvaldi J TI - Monensin and tunicamycin-induced inhibition of HT29 cell spreading and growth. AB - HT29 cells originating from a human colon adenocarcinoma, spread very rapidly after seeding on their own extracellular matrix (ECM). Preincubation of cells with the inhibitor of protein glycosylation tunicamycin (TM) or with the ionophore monensin substantially suppressed cell spreading in serum-free medium without affecting cell adhesion to ECM. Addition of the drugs after cell attachment and spreading inhibited cell growth. TM-treated cells remained viable after 6 days of exposure to 2 micrograms ml-1 of TM and resumed their normal growth rate and shape after removing the drug from the medium. On the contrary, monensin inhibition of cell growth was not reversible: after 3 days, cells detached from the ECM and were unable to exclude Trypan Blue. At the ultrastructural level, a swollen Golgi apparatus with numerous vacuoles was observed after treatment for 2 h in either TM or monensin-preincubated cells. These results suggest that TM and monensin interfere with the insertion and, or, function of one or more cell surface glycoproteins, possibly interacting with cytoskeleton and involved in cell spreading and growth. MH - Adenocarcinoma/*PATHOLOGY ; Cell Adhesion/DRUG EFFECTS ; Cell Line ; Colonic Neoplasms/*PATHOLOGY ; Furans/*PHARMACODYNAMICS ; Glucosamine/ *ANALOGS & DERIVATIVES ; Human ; Microscopy, Electron ; Monensin/ *PHARMACODYNAMICS ; Support, Non-U.S. Gov't ; Tunicamycin/ *PHARMACODYNAMICS SO - J Cell Sci 1986 Feb;80:269-80 33 UI - 86250251 AU - Arundel CM ; Kenney SM ; Leith JT ; Glicksman AS TI - Contrasting effects of the differentiating agent sodium butyrate on recovery processes after x-irradiation in heterogeneous human colon tumor cells. AB - The X ray survival responses of two clonal subpopulations of cells (clones A and D) from a heterogeneous human colon adenocarcinoma (DLD-l) were studied in unfed plateau phase cultures either as control cultures or in cultures grown for three passages in medium containing the differentiating agent sodium butyrate (NaB, 2 mM). Specifically, the cultures were studied with regard to their ability to express both potentially lethal and sublethal damage recovery (PLDR and SLDR). Growth in NaB-containing medium enhanced the radiation sensitivity of both cell lines in the low dose ("shoulder:) region of the survival curve. For clone A, the Dq value was reduced by 59%, and for clone D, the Dq value was reduced by 96%. NaB treatment increased both the rate and the extent of PLDR in both cell lines as assessed by single dose kinetic studies. However, when split dose experiments are performed to assess the expression of SLDR, NaB pretreatment was shown to totally inhibit the expression of SLDR, and also to alter the expression of PLDR under these conditions. These data suggest that PLDR and SLDR are separate, yet related, cellular recovery processes. In addition, NaB may be useful as an adjunct to radiotherapy by virtue of its ability to sensitize tumor cells in the dose range conventionally used for therapy, as well as by inhibition of sublethal damage recovery. MH - Adenocarcinoma/PATHOLOGY/*RADIOTHERAPY ; Butyrates/*PHARMACODYNAMICS ; Butyric Acids/*PHARMACODYNAMICS ; Cell Differentiation/DRUG EFFECTS ; Cell Division/DRUG EFFECTS/RADIATION EFFECTS ; Cell Line ; Cell Survival/ DRUG EFFECTS/RADIATION EFFECTS ; Clone Cells ; Colonic Neoplasms/ PATHOLOGY/*RADIOTHERAPY ; Dose-Response Relationship, Radiation ; Human ; Kinetics ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Int J Radiat Oncol Biol Phys 1986 Jun;12(6):959-68 34 UI - 86243281 AU - Fukushi Y ; Nudelman E ; Levery SB ; Higuchi T ; Hakomori S TI - A novel disialoganglioside (IV3NeuAcIII6NeuAcLc4) of human adenocarcinoma and the monoclonal antibody (FH9) defining this disialosyl structure. AB - This ganglioside is highly immunogenic, and immunization of mice with this disialoganglioside fraction coated on Salmonella minnesota followed by fusion of immunized spleen cells with mouse myeloma and selection of the hybridoma by positive reactivity with the purified disialoganglioside resulted in the establishment of a hybridoma secreting immunoglobulin G2a antibody FH9 that reacts specifically with the ganglioside antigen above but not with monosialosyllactotetraosylceramide I (IV3NeuAcLc4), monosialosyllactotetraosylceramide II (III6NeuAcLc4), or any other gangliosides tested. MH - Adenocarcinoma/*ANALYSIS ; Antibodies, Monoclonal ; Carbohydrate Conformation ; Carbohydrate Sequence ; Cell Line ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Colonic Neoplasms/ *ANALYSIS ; Erythrocyte Membrane/ANALYSIS ; Gangliosides/IMMUNOLOGY/ *ISOLATION & PURIFICATION ; Human ; Liver Neoplasms/ANALYSIS/*SECONDARY ; Sialic Acids/ANALYSIS ; Spectrum Analysis, Mass ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Biochemistry 1986 May 20;25(10):2859-66 35 UI - 86243035 AU - Kirkland SC ; Bailey IG TI - Establishment and characterisation of six human colorectal adenocarcinoma cell lines. AB - The establishment and characterisation (morphology, ultrastructure, tumourigenicity) of six cell lines from primary human colorectal adenocarcinomas is described. These lines were established from surgical specimens, from 49 unselected patients, without the use of 'feeder' cells, 'conditioned' medium or passage of cells in nude mice. The six cell lines exhibit considerable variation in morphology, CEA secretion and tumourigenicity in nude mice. At least two of the lines retain some of the differentiated characteristics of colorectal epithelium. MH - Adenocarcinoma/*PATHOLOGY/ULTRASTRUCTURE ; Aged ; Animal ; Carcinoembryonic Antigen/ANALYSIS ; *Cell Line ; Colonic Neoplasms/ *PATHOLOGY/ULTRASTRUCTURE ; Female ; Human ; Male ; Mice ; Mice, Nude ; Middle Age ; Neoplasm Transplantation ; Rectal Neoplasms/*PATHOLOGY/ ULTRASTRUCTURE ; Support, Non-U.S. Gov't ; Transplantation, Heterologous SO - Br J Cancer 1986 Jun;53(6):779-85 36 UI - 86160929 AU - Tan MH ; Nowak NJ ; Loor R ; Ochi H ; Sandberg AA ; Lopez C ; Pickren JW ; Berjian R ; Douglass HO Jr ; Chu TM TI - Characterization of a new primary human pancreatic tumor line. AB - A primary human pancreatic tumor line (BxPC-3) has been established from a biopsy specimen of a histologically confirmed adenocarcinoma of the body of the pancreas. Tumorigenicity was proven by xenograft in athymic nude mice. Upon re-establishment of tumor xenografts in tissue culture, the epithelial tumor cells retained their original morphology. Histopathologically, the tumors grown in nude mice exhibited the original characteristics of the primary adenocarcinoma in the patient, producing traceable mucin and displaying moderately well to poorly differentiated adenocarcinomas with occasional lymphocytic infiltrations at the tumor peripheries. Furthermore, the tumor xenografts differentially expressed carcinoembryonic antigen, human pancreas cancer-associated antigen, and human pancreas-specific antigen. Karyotyping and glucose-6-phosphate dehydrogenase isoenzyme characterization revealed that this tumor line was of human origin and devoid of HeLa cell contamination. The BxPC-3 tumor line has been maintained for more than four years in our laboratory and represents a valuable model for primary human pancreatic cancer. MH - Adenocarcinoma/FAMILIAL & GENETIC/*PATHOLOGY ; Animal ; Biopsy ; Cell Line ; Chromosome Banding ; Human ; Karyotyping ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Pancreatic Neoplasms/FAMILIAL & GENETIC/ *PATHOLOGY ; Support, U.S. Gov't, P.H.S. ; Tissue Culture/METHODS ; Transplantation, Heterologous SO - Cancer Invest 1986;4(1):15-23 37 UI - 86053225 AU - Phillips JL ; Rutledge L ; Winters WD TI - Transferrin binding to two human colon carcinoma cell lines: characterization and effect of 60-Hz electromagnetic fields. AB - 125I-Labeled human transferrin was used to study the binding of transferrin to Colo 320 DM and Colo 205 human cell lines derived from adenocarcinomas of the colon. Although transferrin uptake was greater in both cases at 37 degrees than at 4 degrees it was found that slightly greater than two-thirds of the transferrin associated with the cells at 37 degrees was not bound to surface receptors but rather had been internalized by the cells. Subsequent analysis of true surface binding at 4 degrees by Scatchard analysis allowed determination of the number of transferrin receptors as well as association constants for the interaction. The number of transferrin receptors per cell was found to be inversely related to the cell density of the cultures from which cells were removed for study. Association constants were unaffected by cell density, with average values of 1.2 and 5.4 X 10(8) M-1 obtained for Colo 320 DM and Colo 205, respectively. Additionally, maximum theoretical numbers of receptors of 1.05 X 10(5)/cell for Colo 320 DM and 1.39 X 10(5)/cell for Colo 205 were determined. Furthermore, exposure of Colo 205 cells to three different experimental situations, i.e., 60 Hz-generated electric field only (E+, 300 mA/m2rms), magnetic field only (M+, 1.0 gauss rms), and combined electric + magnetic fields at these intensities (E+M+), altered the expression of transferrin receptors as compared to a concurrently run unexposed control population of cells (E-M-). In three separate experiments the number of transferrin receptors quantitated on both M+ and E+M+ cells was independent of cell culture density and was close to or exceeded the maximum theoretical number of receptors determined for this cell line. In contrast, E+ cells expressed fewer transferrin receptors than was predicted on the basis of cell culture density. MH - Adenocarcinoma/*METABOLISM ; Cell Line ; Cell Membrane/PHYSIOLOGY ; Colonic Neoplasms/*METABOLISM ; Electromagnetics ; Human ; Kinetics ; Receptors, Endogenous Substances/*METABOLISM ; Support, Non-U.S. Gov't ; Temperature ; Transferrin/*METABOLISM SO - Cancer Res 1986 Jan;46(1):239-44 38 UI - 86215957 AU - Twentyman PR ; Fox NE ; Wright KA ; Bleehen NM TI - Derivation and preliminary characterisation of adriamycin resistant lines of human lung cancer cells. AB - We have produced adriamycin (ADM)-resistant variants of the human lung cancer cell lines NCI-H69 (small cell), MOR (adenocarcinoma) and COR-L23 (large cell) but have failed to produce resistant variants of two other small cell lines. In each case, the derivation protocol took 7-9 months and included a period of drug-free growth. All three resistant lines show reduced cellular content of ADM after 1 h exposure when compared with their controls. During prolonged incubation of control and resistant NCI-H69 cells in 0.4 microgram ml-1 ADM, the ADM content of resistant cells was 6-7 times lower than that of control cells. The ratio of ADM doses to suppress growth of the two lines, however, was in the range of 40-200X. The ADM-resistant variant of NCI-H69 was also resistant to vincristine, colchicine, VP16, mitozantrone, 4' epiadriamycin and 4' deoxyadriamycin, somewhat resistant to melphalan but not resistant to aclacinomycin A, bleomycin of CCNU. The resistance to ADM could be partially overcome by the use of verapamil, an inhibitor of calcium transport. MH - Adenocarcinoma/*PATHOLOGY ; Carcinoma, Oat Cell/*PATHOLOGY ; Cell Line ; Cytological Technics ; Doxorubicin/ANALYSIS/*PHARMACODYNAMICS ; Drug Resistance ; Human ; Lung Neoplasms/*PATHOLOGY ; Tumor Stem Cell Assay ; Verapamil/PHARMACODYNAMICS SO - Br J Cancer 1986 Apr;53(4):529-37 39 UI - 86215422 AU - Dunzendorfer U ; Balis ME ; Whitmore WF TI - Some aspects of clearance of mitoguazone in cancer patients and experimental cancer models. AB - Mitoguazone (methylglyoxal-bis(guanyl-hydrazone), MGBG) was studied by its first-pass mechanism in both cancer patients and experimental cancer models. It appears from the study that 90% of MGBG is cleared from the plasma within minutes. 24-h recovery in the urine, however, did not exceed 16% so that 84% of the drug seems to be bound to subcellular compartments. Tissue levels of MGBG in the normal prostate ranged higher than in experimental prostate cancer type 3327 M/G, i.e. enhanced clearance from cancer tissues: polyamine biosynthetic enzymes ornithine decarboxylase as well as S-adenosylmethionine decarboxylase are contrarily affected by MGBG. MH - Adenocarcinoma/*DRUG THERAPY ; Animal ; Carcinoma, Renal Cell/*DRUG THERAPY ; Cell Line ; Cells, Cultured ; Clinical Trials ; Dose-Response Relationship, Drug ; Drug Administration Schedule ; Human ; Infusions, Parenteral ; Kidney Neoplasms/*DRUG THERAPY ; Male ; Mice ; Mice, Inbred Strains ; Mitoguazone/ADMINISTRATION & DOSAGE/*METABOLISM/TOXICITY ; Ornithine Decarboxylase/ANALYSIS ; Prostatic Neoplasms/*DRUG THERAPY ; Rats ; Rats, Inbred Strains ; S-Adenosylmethionine Decarboxylase/ ANTAGONISTS & INHIBITORS/ANALYSIS ; Support, Non-U.S. Gov't ; Tissue Distribution SO - Arzneimittelforschung 1986 Mar;36(3):506-8 40 UI - 86211657 AU - Cheret AM ; Laboisse CL ; Roumagnac I ; Augeron C ; Lewin MJ TI - Highly histamine-responsive clones from the human gastric adenocarcinoma cell line HGT-1. AB - The human gastric epithelial cell line HGT-1 possesses adenylate cyclase-coupled histamine H2 receptors. To test the cellular homogeneity or heterogeneity with respect to these receptors, we have isolated 7 clones from the HGT-1 line and studied their basal and histamine-stimulated adenylate cyclase activities. Basal adenylate cyclase activities of the clones did not differ significantly, nor did 10 mM NaF- nor 0.1 mM Gpp(NH)p-stimulated activities. However, histamine stimulation of adenylate cyclase varied among clones from 1.9 fold to 5.4 fold basal activity. The EC50 values, determined in 3 clones, were not significantly different. These findings support the heterogeneity of histamine responsiveness of the human gastric cell line HGT-1. In addition, they suggest that highly histamine-responsive clones may be useful models to study the gastric histamine H2-receptor and its specific antagonists in the human. MH - Adenocarcinoma/*ENZYMOLOGY ; Adenyl Cyclase/*METABOLISM ; Cell Line ; Cimetidine/PHARMACODYNAMICS ; Clone Cells ; Diphenhydramine/ PHARMACODYNAMICS ; Guanylyl Imidodiphosphate/PHARMACODYNAMICS ; Histamine/ *PHARMACODYNAMICS ; Human ; Kinetics ; Sodium Fluoride/PHARMACODYNAMICS ; Stomach Neoplasms/*ENZYMOLOGY ; Support, Non-U.S. Gov't SO - Agents Actions 1986 Mar;17(5-6):436-40 41 UI - 86201752 AU - Holinka CF ; Hata H ; Kuramoto H ; Gurpide E TI - Responses to estradiol in a human endometrial adenocarcinoma cell line (Ishikawa). AB - A human endometrial adenocarcinoma cell line (Ishikawa) has been found to be estrogen responsive. The growth stimulatory effects of estradiol (10(-8) M) could be clearly demonstrated when cell cultures containing the hormone were compared with the maximal cell density achieved in control cultures. The approx. 3-fold increase in cell density observed 2-3 weeks after plating, with frequent medium changes, could by blocked by a 100-fold molar excess of the antiestrogen trans-4-monohydroxytamoxifen. When added to hormone-free cultures that had reached a plateau level of cell numbers on day 14 after plating, estradiol (10(-8) M) caused the resumption of proliferation: after 6 days in the presence of the hormone, the cultures contained nearly twice the cell numbers of controls. Effects of estradiol on Ishikawa cells were also evident from the several-fold increases in the levels of specific progesterone binders provoked by the hormone at 10(-9)-10(-6) M concentrations. Cells injected into nude mice formed tumors which contained estrogen and progesterone binders. The availability of a fast-growing (doubling time approx. 30 h) endometrial cancer cell line responsive to estradiol at near physiologic levels will facilitate biochemical studies of hormonal effects on the human endometrium. MH - Adenocarcinoma/METABOLISM/*PATHOLOGY ; Animal ; Cell Division/DRUG EFFECTS ; Cell Line ; Estradiol/*PHARMACODYNAMICS ; Female ; Human ; Mice ; Mice, Nude ; Progesterone/METABOLISM ; Receptors, Estrogen/ANALYSIS ; Support, U.S. Gov't, P.H.S. ; Uterine Neoplasms/METABOLISM/*PATHOLOGY SO - J Steroid Biochem 1986 Jan;24(1):85-9 42 UI - 86195531 AU - Lee YC ; Saijo N ; Sasaki Y ; Takahashi H ; Sakurai M ; Ishihara J ; Sano T ; Hoshi A ; Chen KM ; Hamburger AW TI - Antitumor effect of two-drug simultaneous or sequential use of cisplatin, vindesine or etoposide on human pulmonary adenocarcinoma cell lines in tumor clonogenic assay. AB - The antitumor effects of two-drug simultaneous or sequential use of cisplatin, vindesine or etoposide were examined in human pulmonary adenocarcinoma cell lines by human tumor clonogenic assay. Different tumor cell lines (PC-9, PC-13 and PC-14) that originated from the same histological cell type responded quite variably to simultaneous and/or sequential therapy. The antitumor effects of various sequential combinations of cisplatin, vindesine and etoposide depended strongly on the nature of the individual tumor cell line tested. MH - Adenocarcinoma/*DRUG THERAPY ; Antineoplastic Agents, Combined/ *THERAPEUTIC USE ; Cell Line ; Cisplatin/ADMINISTRATION & DOSAGE ; *Colony-Forming Units Assay ; Etoposide/ADMINISTRATION & DOSAGE ; Human ; Lung Neoplasms/*DRUG THERAPY ; Support, Non-U.S. Gov't ; *Tumor Stem Cell Assay ; Vindesine/ADMINISTRATION & DOSAGE SO - Jpn J Cancer Res 1986 Mar;77(3):312-8 43 UI - 86186933 AU - Houghton JA ; Weiss KD ; Williams LG ; Torrance PM ; Houghton PJ TI - Relationship between 5-fluoro-2'-deoxyuridylate, 2'-deoxyuridylate, and thymidylate synthase activity subsequent to 5-fluorouracil administration, in xenografts of human colon adenocarcinomas. AB - 5-Fluorouracil (FUra) has been administered to mice bearing xenografts of human colon adenocarcinomas. In two tumor lines, HxGC3 and HxVRC5, intrinsically resistant to FUra, 2'-deoxyuridylate (dUMP) accumulated 13.4- and 23.9-fold above basal levels. In HxELC2 xenografts, which demonstrated some sensitivity to FUra, there was a decrease in dUMP concentration after drug administration. Maximal intratumor levels of 5-fluoro-2'-deoxyuridylate (FdUMP) were found at 1 hr, but decreased in all tumor lines by 4 hr after administration of FUra. Data derived in tumor cytosols suggested that FdUMP levels in situ were not rate-limiting for formation of covalent ternary complex, but that accumulation of dUMP would retard the rate of complex formation. Subsequent to administration of FUra, thymidylate synthase activity was reduced greater than 75% in all tumors, but it recovered rapidly in tumors resistant to FUra. In addition, the pretreatment level of activity of thymidylate synthase was 12.7-fold greater in HxVRC5 tumors than in HxELC2 tumors. This elevated activity in HxVRC5 tumors appears not to be a consequence of gene amplification. Formation of FdUMP or the accumulation of dUMP did not correlate with the activity of phosphatases measured at pH 5.8 or pH 9.2 in each tumor line. Further, inhibition of phosphatase activity did not alter, significantly, the net rate of dissociation of the FdUMP-thymidylate synthase-[6R]-CH2-H4PteGlu complex. MH - Adenocarcinoma/*METABOLISM ; Adenosine Monophosphate/PHARMACODYNAMICS ; Animal ; Cell Line ; Colonic Neoplasms/*METABOLISM ; Deoxyuracil Nucleotides/ANALYSIS/*METABOLISM ; Drug Resistance ; Fluorodeoxyuridylate/ ANALYSIS/*METABOLISM ; Fluorouracil/*PHARMACODYNAMICS ; Human ; Hydrogen-Ion Concentration ; Mice ; Neoplasm Transplantation ; Phosphatases/ANALYSIS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Thymidylate Synthetase/*ANALYSIS ; Transplantation, Heterologous SO - Biochem Pharmacol 1986 Apr 15;35(8):1351-8 44 UI - 86182910 AU - Motoyama T ; Hojo H ; Watanabe H TI - Comparison of seven cell lines derived from human gastric carcinomas. AB - In an attempt to elucidate various histological features of gastric cancers, seven human gastric adenocarcinomas were studied in vitro and in nude mice. Growth pattern of each cultured cell line in vitro corresponded well to the histological type of parent tumor. The cell lines, MKN7, MKN74, and MKN28 derived from differentiated carcinomas showed morphological characteristics of intestinal differentiation in cell polarity and microvilli with core-filaments in vitro as well as in nude mice. However, they gradually diminished the characteristics in course of time. The cell lines, MKN 45 and OKAJIMA, derived from undifferentiated carcinomas, had natures of not only ordinary gastric mucosa but also intestinal metaplastic mucosa. They seem to have multipotentiality for differentiation, and preserved well the natures for long periods of culture. The KWS-I cell line composed of undifferentiated cells in vitro displayed the potential for differentiation in nude mice. However, the differentiation of KATO-III cells derived from a signet-ring cell carcinoma was suppressed in nude mice. The common abnormality of chromosome was not found, and the growth rate in vitro was not dependent on the histological type of parent tumor. MH - Adenocarcinoma/*PATHOLOGY/ULTRASTRUCTURE ; Animal ; Cell Line ; Comparative Study ; Human ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron ; Neoplasm Transplantation ; Stomach Neoplasms/*PATHOLOGY/ ULTRASTRUCTURE ; Support, Non-U.S. Gov't ; Tissue Culture/METHODS ; Transplantation, Heterologous SO - Acta Pathol Jpn 1986 Jan;36(1):65-83 45 UI - 86177571 AU - Tsunokawa Y ; Takebe N ; Kasamatsu T ; Terada M ; Sugimura T TI - Transforming activity of human papillomavirus type 16 DNA sequence in a cervical cancer. AB - A genomic DNA sample from cervical cancer tissue, containing human papillomavirus (HPV) type 16, was found to induce malignant transformation of NIH 3T3 cells when it was tested by transfection assays using the calcium phosphate coprecipitation technique. The primary and secondary transformants contained the HPV type 16 DNA sequences and human specific Alu family sequences. To the best of our knowledge, it has not been reported previously that HPV type 16 DNA sequences in total genomic DNA from a cervical cancer have transforming activity. MH - Adenocarcinoma/*FAMILIAL & GENETIC/MICROBIOLOGY ; Animal ; Cell Line ; *Cell Transformation, Viral ; Cervix Neoplasms/*FAMILIAL & GENETIC/ MICROBIOLOGY ; DNA, Neoplasm/*GENETICS ; DNA, Viral/*GENETICS ; Female ; Human ; Mice ; Papillomaviruses/*GENETICS ; Repetitive Sequences, Nucleic Acid ; Support, Non-U.S. Gov't SO - Proc Natl Acad Sci USA 1986 Apr;83(7):2200-3 46 UI - 86168210 AU - Gowda DC ; Bhavanandan VP ; Davidson EA TI - Structures of O-linked oligosaccharides present in the proteoglycans secreted by human mammary epithelial cells. AB - The structures of O-glycosidically linked oligosaccharides present in the heparan sulfate and chondroitin sulfate proteoglycans isolated from the culture medium of a normal (HBL-100) and a malignant (MDA-MB-231) human mammary epithelial cell line have been determined. Both proteoglycan types from the two cell lines contain a series of O-linked oligosaccharides ranging in size from di- to hexasaccharide. Cells were grown in the presence of either [3H]glucosamine or [3H]galactose and Na2 35SO4, and the proteoglycans were isolated as described (Gowda, D. C., Bhavanandan, V. P., and Davidson, E. A. (1986) J. Biol. Chem. 261, 4926-4934). The O-linked oligosaccharides were released from the proteoglycans by alkaline borohydride treatment and purified by a combination of gel filtration and high voltage paper electrophoresis. The structures of two neutral and seven acidic oligosaccharides were established based on sugar composition, the results of periodate oxidation, sequential exoglycosidase treatment, and methylation analysis. Periodate oxidation, taking advantage of tritium label at specific positions of constituent sugars, proved to be a valuable tool in establishing the structure of isomeric components in the mixture. The structures of the oligosaccharides were assigned as follows: (Formula: see text) The oligosaccharide containing both sialic acid and ester sulfate is novel and has not been reported previously. MH - Adenocarcinoma/*SECRETION ; Borohydrides ; Breast/*SECRETION ; Breast Neoplasms/*SECRETION ; Carbohydrate Conformation ; Cell Line ; Chromatography, Gel ; Electrophoresis, Paper ; Epithelium/SECRETION ; Female ; Glycoside Hydrolases/PHARMACODYNAMICS ; Human ; Indicators and Reagents ; Methylation ; Oligosaccharides/*ISOLATION & PURIFICATION ; Oxidation-Reduction ; Periodic Acids ; Proteoglycans/ANALYSIS/*SECRETION ; Support, U.S. Gov't, P.H.S. SO - J Biol Chem 1986 Apr 15;261(11):4935-9 47 UI - 86168209 AU - Gowda DC ; Bhavanandan VP ; Davidson EA TI - Isolation and characterization of proteoglycans secreted by normal and malignant human mammary epithelial cells. AB - The proteoglycans secreted by a malignant human breast cell line (MDA-MB-231) were compared with the corresponding proteoglycans from a normal human breast cell line (HBL-100). The physicochemical characteristics of these proteoglycans were established by hexosamine analysis, chemical and enzymatic degradations, and dissociative cesium chloride density gradient centrifugation, and by gel filtration before and after alkaline beta-elimination. Both cell lines secreted approximately 70% of the synthesized proteoglycans, which were composed of 20% heparan sulfate and 80% chondroitin sulfate proteoglycans. The MDA cell line secreted large hydrodynamic size (major) and small hydrodynamic size heparan sulfate proteoglycan. In contrast HBL cells secreted only one species having a hydrodynamic size intermediate to the above two. The chondroitin sulfate proteoglycans from MDA medium were slightly larger than the corresponding polymers from HBL medium. All proteoglycans except the small hydrodynamic size heparan sulfate proteoglycan from MDA medium were of high buoyant density. The proteoglycans of both cell lines contained significant proportions of disulfide-linked lower molecular weight components which were more pronounced in the proteoheparan sulfate polymers, particularly those from MDA medium, than in chondroitin sulfate proteoglycans. The glycosaminoglycans of heparan sulfate proteoglycans from MDA medium were more heterogeneous than those from HBL medium. The glycosaminoglycan chains of large hydrodynamic size heparan sulfate proteoglycans from MDA medium were larger in size than those from HBL medium while small hydrodynamic size heparan sulfate proteoglycans contained shorter glycosaminoglycan chains. In contrast to the glycosaminoglycans derived from chondroitin sulfate proteoglycans of both MDA and HBL medium were comparable in size. The heparan sulfate as well as chondroitin sulfate proteoglycans of both cell lines contained both neutral (di- and tetrasaccharides) and sialylated (tri- to hexasaccharides) O-linked oligosaccharides. MH - Adenocarcinoma/*METABOLISM ; Breast/*SECRETION ; Breast Neoplasms/ *SECRETION ; Carbohydrate Conformation ; Cell Line ; Centrifugation, Density Gradient ; Chondroitin Sulfates/ANALYSIS ; Chromatography, Gel ; Disulfides/ANALYSIS ; Epithelium/SECRETION ; Female ; Galactosamine/ ANALYSIS ; Glucosamine/ANALYSIS ; Glycosaminoglycans/ANALYSIS ; Heparitin Sulfate/ANALYSIS/METABOLISM ; Human ; Molecular Weight ; Oligosaccharides/ ANALYSIS ; Proteoglycans/ANALYSIS/*SECRETION ; Support, U.S. Gov't, P.H.S. SO - J Biol Chem 1986 Apr 15;261(11):4926-34 48 UI - 86165283 AU - Heckl W ; Fogh J TI - Human lactic dehydrogenase as a marker for monitoring the growth of prostatic carcinoma in nude mice. AB - Human lactate dehydrogenase (1,1,1,27-LDH) isoenzymes were determined in human prostatic carcinoma cell lines. The isoenzymes LDH-4/LDH-5 were identified and quantified in the serum of athymic mice bearing the human prostatic carcinoma cell lines PC-3, PC-3/M and DU 145 subcutaneously. The amounts of the slowly migrating isoenzymes released by the solid tumors correlated with increasing tumor volume and tumor weight. These correlations indicate that the human LDH-4/LDH-5 isoenzymes are useful parameters for monitoring the presence and growth of prostatic cancer in the athymic animal model. MH - Adenocarcinoma/*ENZYMOLOGY/PATHOLOGY ; Animal ; Cell Line ; Disease Models, Animal ; Female ; Human ; Lactate Dehydrogenase Isoenzymes/ *ANALYSIS ; Male ; Mice ; Mice, Nude ; Prostatic Neoplasms/*ENZYMOLOGY/ PATHOLOGY SO - Exp Cell Biol 1986;54(1):34-9 49 UI - 86163519 AU - Gazdar AF TI - Advances in the biology of non-small cell lung cancer. AB - We have initiated an in-depth, comprehensive study of non-SCLC in the expectation that improvements in clinical management and diagnosis of lung cancer patients will follow. We have established and characterized over 30 such tumors in long-term culture and as xenografts in athymic nude mice. These models usually retain the morphologic and other properties of the tumors from which they were derived. With the use of fully defined medium, most SCLC and adenocarcinomas can be established as long-term cultures. The growth conditions for squamous cell carcinomas have not been fully defined, and the success rate is low. In contrast to SCLC, there is considerable heterogeneity of non-SCLC tumors, with more than 12 phenotypes identified among the first 30 lines established. In addition, there is considerable overlap of properties, both within the non-SCLC types as well as between non-SCLC and SCLC. These findings indicate a common origin for all lung cancers. Radiation and drug sensitivity studies suggest that cell line data may correlate with clinical responses. Clinical protocols utilizing our ability to culture and drug test individual tumors are currently under way. They represent the first steps in a combined clinico-laboratory approach to the management of lung cancer. MH - Adenocarcinoma/CLASSIFICATION/*PATHOLOGY ; Animal ; Carcinoma, Oat Cell/ CLASSIFICATION/PATHOLOGY ; Carcinoma, Squamous Cell/CLASSIFICATION/ *PATHOLOGY ; Cell Division ; Cell Line ; DNA, Neoplasm/ANALYSIS ; Epidermal Growth Factor-Urogastrone/METABOLISM ; Human ; Lung Neoplasms/ CLASSIFICATION/*PATHOLOGY ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Ploidies ; Transplantation, Heterologous SO - Chest 1986 Apr;89(4 Suppl):277S-283S 50 UI - 86116238 AU - Braakhuis BJ ; Nauta MM ; Romijn JC ; Rutgers DH ; Smink T TI - Enhanced success rate of transplantation with human tumors in cyclophosphamide-treated nude mice. AB - Immunosuppression was investigated to enhance the take rate of various human tumor types in nude BALB/c or B10.LP/Cpb mice. Cyclophosphamide (CY), known as an immunosuppressive agent, was injected ip at a dose of 100 mg/kg 24 hours prior to subcutaneous implantation of tumor fragments obtained from established xenograft lines or from patients. In 3 out of 8 ovarian adenocarcinoma tumor lines, with a take of 50% or less in untreated control animals, tumor take was significantly increased by CY treatment to values between 75 and 100%. No effect of CY treatment on tumor take was seen with squamous cell carcinomas of the head and neck region and with lung and prostatic carcinomas. The growth rate of these xenografts was not affected by CY pretreatment. These results indicate that immunologic mechanisms indeed can play a role in the inhibition of tumor growth in nude mice. CY pretreatment may enhance the success rate of transplantation, but this effect appears to be limited to certain tumors. MH - Adenocarcinoma/*IMMUNOLOGY/PATHOLOGY ; Animal ; Cell Division ; Cell Line ; Comparative Study ; Cyclophosphamide/ADMINISTRATION & DOSAGE/ *IMMUNOLOGY ; Female ; Human ; Killer Cells, Natural/DRUG EFFECTS/ IMMUNOLOGY ; Male ; Mice ; Mice, Inbred Strains ; Mice, Nude ; Ovarian Neoplasms/*IMMUNOLOGY/PATHOLOGY ; Support, Non-U.S. Gov't ; Transplantation Immunology SO - JNCI 1986 Feb;76(2):241-5 51 UI - 86106576 AU - Falzon M ; McMahon JB ; Gazdar AF ; Schuller HM TI - Preferential metabolism of N-nitrosodiethylamine by two cell lines derived from human pulmonary adenocarcinomas. AB - Diethylnitrosamine (DEN), in common with other nitrosamines, is a carcinogenic agent which produces tumors in a wide variety of tissues in experimental animals. The pulmonary Clara cell is a major target of N-nitrosamine-induced carcinogenesis in hamsters and rats. DEN is believed to require metabolic activation to elicit its carcinogenic effects. The metabolism of [14C]DEN was studied in two cell lines derived from human lung adenocarcinomas and two cell lines derived from human small cell lung cancers by monitoring 14CO2 production and covalent binding of radiolabel from [14C]DEN to the cell protein and DNA fractions. [14C]DEN was metabolized by adenocarcinoma-derived NCI-H322 (with Clara cell features) and NCI-H358 (with features of alveolar type II cells) but not by NCI-H69 and NCI-H128 (derived from small cell carcinoma). Metabolism was markedly inhibited by heat denaturation of the cell protein. [14C]DEN metabolism by NCI-H322 was greatly decreased when the incubation was carried out under anaerobic conditions and in the presence of a carbon monoxide enriched atmosphere. These results suggested the involvement of the cytochrome P-450-dependent monooxygenase enzyme system. Metabolism by NCI-H358 was also decreased in the absence of oxygen or presence of carbon monoxide although the effects were relatively small compared with the results with NCI-H322. On the other hand, aspirin or indomethacin, which are inhibitors of the fatty acid cyclooxygenase component of prostaglandin endoperoxide synthetase, preferentially inhibited [14C]DEN metabolism by NIC-H358. There were little or no effects of these inhibitors on the metabolism of DEN in NCI-H322. The data suggest that DEN metabolism in different lung cell types may be carried out by different enzyme systems which in turn may contribute to the selective effect of DEN in the lung. MH - Adenocarcinoma/*METABOLISM ; Carbon Dioxide/ANALYSIS ; Carbon Radioisotopes ; Cell Line ; Diethylnitrosamine/*METABOLISM ; Human ; Kinetics ; Lung Neoplasms/*METABOLISM ; Radioisotope Dilution Technic SO - Carcinogenesis 1986 Jan;7(1):17-22 52 UI - 86105707 AU - Yoshida H ; Azuma M ; Yanagawa T ; Yura Y ; Hayashi Y ; Sato M TI - Effect of dibutyryl cyclic AMP on morphologic features and biologic markers of a human salivary gland adenocarcinoma cell line in culture. AB - Dibutyryl cyclic AMP (dB-cAMP) has marked effects on the growth, morphologic features, and biologic markers of a human salivary gland adenocarcinoma cell line in culture. A cell line with ultrastructure and biologic markers specific to the intercalated duct cells of human salivary glands was cultivated in the presence of dB-cAMP. Major alterations, such as process formation and expression of myofilaments and oxytocin receptor in addition to myosin and S-100 protein, were observed in those cells with a phenotype similar to myoepithelial cells. Both the anchorage-independent and anchorage-dependent growths were markedly suppressed in the presence of dB-cAMP. After the removal of dB-cAMP from the culture, the treated cells returned rapidly to the phenotype of the untreated cells. These findings indicate that reversible differentiation into the myoepithelial cells of a human salivary gland adenocarcinoma cell line occurs in growth medium containing dB-cAMP. MH - Adenocarcinoma/ANALYSIS/*PATHOLOGY ; Autoradiography ; Carcinoembryonic Antigen/ANALYSIS ; Cell Cycle ; Cell Line ; Clone Cells ; Culture Media ; Dibutyryl Cyclic AMP/*PHARMACODYNAMICS ; Histocytochemistry ; Human ; Immunoenzyme Technics ; Microscopy, Electron ; Myosin/ANALYSIS ; Nerve Tissue Protein S 100/ANALYSIS ; Receptors, Angiotensin/ANALYSIS ; Salivary Gland Neoplasms/ANALYSIS/*PATHOLOGY ; Support, Non-U.S. Gov't SO - Cancer 1986 Mar 1;57(5):1011-8 53 UI - 86079177 AU - Brower M ; Carney DN ; Oie HK ; Gazdar AF ; Minna JD TI - Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serum-free defined medium. AB - We tested the ability of serum-free media to support the in vitro growth of human non-small cell lung carcinoma. A medium containing insulin, transferrin, sodium selenite, hydrocortisone, epidermal growth factor, and bovine serum albumin (1 mg/ml) with serum precoating of culture dishes (modified LA medium) supported three previously established cell lines of non-small cell lung cancer and prevented fibroblast proliferation in fresh tumor specimens but did not support long term tumor cell growth from fresh specimens. We added triiodothyronine, sodium pyruvate, and additional glutamine, insulin, and epidermal growth factor to modified LA medium, precoated with fibronectin and collagen instead of serum, and deleted bovine serum albumin, defining a new medium called ACL-3. ACL-3 medium alone supported the short term growth of 10 of 12 cell lines and the soft agarose cloning of 9 of 12 cell lines tested, and ACL-3 supplemented by an optimal concentration of bovine serum albumin (5 mg/ml) supported the long term growth of 10 of 12 cell lines tested. Moreover, we have grown tumor cells for more than 6 months from 11 of 33 (33%) consecutive fresh clinical specimens of human lung adenocarcinoma in ACL-3 with bovine serum albumin. ACL-3 medium provides a defined environment for the study of growth factor requirements of human non-small cell lung cancer and enhances our ability to grow human lung cancer, particularly adenocarcinoma, in vitro. MH - Adenocarcinoma/*PATHOLOGY ; Carcinoma, Squamous Cell/PATHOLOGY ; Cell Division/DRUG EFFECTS ; Cell Line ; *Culture Media ; Extracellular Matrix/ PHYSIOLOGY ; Growth Substances/PHARMACODYNAMICS ; Human ; Lung Neoplasms/ *PATHOLOGY ; Sepharose ; Tissue Culture SO - Cancer Res 1986 Feb;46(2):798-806