Summary
Established human mammary tumor cell lines constitute an important tool in the study of breast cancer. The aim of this work was to isolate and characterize two new mammary tumor cell lines, JCK and GCS, which were obtained from the pleural effusion and ascitic fluid, respectively, from two breast cancer patients. Both cell lines had some properties of transformed cells, namely immortalization and growth in soft agar. The carcinoma cells presented epithelial morphology shown by light and electron microscopy, and antigenic properties shown by different tumor markers such as a cytokeratin cocktail, carcinoembryonic antigen, epithelial membrane antigen, and human milk fat globule membrane antigen. A significant increase was also found (P>0.05) in cell growth and3H-thymidine incorporation into DNA in the JCK and GCS cell lines in the presence of 17β estradiol at concentrations of 10−9 and 10−7 M, respectively, after 5 days in culture. These cells presented estradiol receptor levels which were similar in the biopsies and the resulting cell lines. The aromatase activity was also similar in the JCK cell line and the original patient biopsy. However, there was a considerably higher aromatase activity in the GCS cell line than in the biopsy specimen. Southern hybridizations with theneu oncogene showed an additional 12 kb fragment in both cell lines, as also seen in patients with breast cancer. We conclude from these studies that thisin vitro system may provide us with a way to study metastatic cells and improve clinical management of breast cancer patients.
References
Hamburger AW, Salmon SE: Primary bioassay of tumor stem cells. Science 197:461–463, 1977
Hamburger A, Salmon SE: Primary bioassay of human myeloma stem cells. J Clin Invest 60:846–854, 1977
Pavelic ZP, Slocum HK, Rustan YM: Growth of cell colonies in soft-agar from biopsies of different human solid tumors. Cancer Res 40:4151–4158, 1980
Brooks SC, Locke ER, Soule HD: Estrogen receptor in human cell line (MCF-7) from carcinoma of breast. J Biol Chem 248:6251–6253, 1973
Cailleau R, Mackay B, Young RK, Reeves WJ Jr: Tissue culture studies on pleural effusions from breast cancer patients. Cancer Res 34:801–809, 1974
Engel LW, Young NA, Tralka TS, Lippman ME, O'Brien SJ, Joice MJ: Establishment and characterization of three new continuous cell lines derived from human breast carcinomas. Cancer Res 38:3352–3364, 1978
Young RK, Cailleau RM, Mackay B, Reeves WJ Jr: Establishment of epithelial cell line MDA-MB-157 from metastatic pleural effusion of human breast carcinoma. In Vitro 9:239–245, 1974
Kohler G, Milstein C: Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497, 1975
Abarca J, Aboud-Pirak E, Otte-Slachmylder C, van Bogaert LJ, Staquet JP, Vancraynes TMP, Schneider YJ, Trouet A: Immunohistochemical screening of 6 monoclonal antibodies to HMFGM and its value in pathology. Path Res Pract 180:249, 1985
Abarca J, Bussalastit G, van Bogaert LJ, Trouet A: Improved diagnosis of carcinoma cells in pleural effusion from patients with breast cancer. 4th EORTC Breast Conference, London, 1987. Prot Biol Fluids 32:553–556, 1985
Ghosh AK, Spriggs AI, Taylor-Papadimitriou J, Mason DY: Immunocytochemical staining of cells in pleural and peritoneal effusions with a panel of monoclonal antibodies. J Clin Pathol 36:1154–1164, 1983
Hilkens J, Buijs F, Hilgers J, Hageman Ph, Calafat J, Sonneberg A, van der Valk M: Monoclonal antibodies against human milk fat globule membrane detecting differentiation antigens. Int J Cancer 34:197–206, 1984
Schlom J, Greiner J, Hand PH, Lolcher D, Inghirami G, Weeks M, Postke S, Fisher PB, Noguchi P, Kufe D: Monoclonal antibodies to breast cancer associated antigens as potential reagents in the management of breast cancer. Cancer 54:2777–2791, 1984
Shively JE, Todd ChW: CEA — Chemistry and Biology in Cancer Markers. Diagnostic and Developmental Significance. Sell S (ed). The Human Press, Clifton, New Jersey, 1980
Walts AE, Said JW, Banks-Schlegel S: Keratin and CEA exfoliated mesothelial and malignant cells: an immunoperoxidase study. Am J Clin Pathol 80: 671–676, 1983
Huseby RA: Steroids and tumorigenesis in experimental animals. In Dorfman RV (ed) Methods in Hormone Research, Vol. 4. Academic Press, New York, 1965, pp 123
Calaf G, Garrido F, Moyano C, Rodriguez R: Influence of hormones on DNA synthesis of breast tumors in culture. Breast Cancer Res Treat 8:223–232, 1986
Chalbos D, Vignon K, Keydar I, Rochefort H: Estrogens stimulate cell proliferation and induce secretory proteins in a human breast cancer cell line (T47D). J Clin Endocr Metab 55:276–283, 1982
Darbre P, Yates J, Curtis S, King RJB: Effects of estradiol on human breast cancer cells in culture. Cancer Res 43:349–354, 1983
Horwitz KB, Costlow ME, McGuire WL: MCF-7, a human breast cancer cell line with estrogen, androgen, progesterone and glucocorticoid receptors. Steroids 26:785–795, 1975
Lippman ME: Hormonal regulation of human breast cancer cells in vitro. Banbury Rep 8:171–181, 1981
Chu MY, Hagerty MG, Wiemann MC, Tibbetts LM, Sato S, Cummings FJ, Bagaars HA, Leduc EH, Calabresi P: Differential characteristics of two newly established human breast carcinoma cell lines. Cancer Res 45:1357–1366, 1985
Keydar I, Chen L, Karby S, Weiss FR, Delerea J, Radu M, Chaiteik S, Brenner HJ: Establishment and characterization of a cell line of human breast carcinoma origin. Eur J Cancer 15:5659–5670, 1979
Lasfargues EJ, Coutinho WG, Redfield ES: Isolation of two human tumor epithelial cell lines from solid breast carcinomas. J Natl Cancer Inst 61:967–978, 1978
Soule HD, Vasquez J, Long A, Albert S, Brennan MJ: A human cell line from a pleural effusion derived from a breast carcinoma. J Natl Cancer Inst 51:1409–1416, 1973
Horwitz KB, Mockus MB, Lessey BA: Variant T-47D human breast cancer cells with high progesterone receptor levels despite estrogen and antiestrogen resistance. Cell 28:633–642, 1982
Yu WCY, Leung BS, Gao YL: Effects of 17β estradiol on progesterone receptors and the uptake of thymidine in human breast cancer cell line CAMA-1. Cancer Res 41:5004–5009, 1981
Allegra JC, Korat O, Do HMT, Lippman ME: The regulation of progesterone receptors by 17 beta estradiol and tamoxifen in the ZR-75-1 human breast cancer cell line in defined medium. J Recept Res 2:17–27, 1981
Welshons WV, Murphy CS, Koch R, Calaf G, Jordan VC: Stimulation of breast cancer cells in vitro by the environmental estrogen enterolactone and the phytoestrogen equol. Breast Cancer Res Treat 10: 169–175, 1987
Horwitz KB, McGuire WL: Antiestrogen mechanism of action and effects in breast cancer. In McGuire WL (ed) Breast Cancer — Advances in Research and Treatment. Plenum, New York, 1978, Vol 2, pp 155–204
Horwitz KB: Is a functional estrogen receptor always required for progesterone receptor induction in breast cancer? J Steroid Biochem 15:209–219, 1981
Horwitz KB, McGuire WL: Estrogen control of progesterone receptor in human breast cancer. Correlation with nuclear processing of estrogen receptor. J Biol Chem 253:2223–2228, 1978
Horwitz KB, Zava DT, Thilagar AK, Jensen EM, McGuire WL: Steroid receptor analysis of nine human breast cancer cell lines. Cancer Res 38:2434–2437, 1978
Lippman ME, Bolan G, Huff K: The effects of estrogens and antiestrogens on hormone responsive human breast cancer in long term tissue culture. Cancer Res 36:4595–4601, 1976
Aitken SC, Lippman MF: Hormonal regulation of de novo pyrimidine synthesis and utilization in human breast cancer cells in tissue culture. Cancer Res 43:4681–4690, 1983
Bates SE, McManaway ME, Lippman ME, Dickson RB: Characterization of estrogen responsive transforming activity in human breast cancer cells. Cancer Res 46:1707–1713, 1986
MacIndoe JH: Estradiol formation from testosterone by continuously cultured human breast cancer cells. J Clin Endocrinol Metab 49:272–277, 1979
Hsueh AJW, Jones PHC, Adashi EY, Wang C, Zhuang L-Z, Welsh TH Jr: Intraovarian mechanisms in the hormonal control of granulosa cell differentiation in rats. Reprod Fertil 69:325–342, 1983
Thompson EA, Siiteri PK: Utilization of oxygen and NADPH by human placental microsomes during aromatization of androstenedione. J Biol Chem 249: 5364–5372, 1974
Coussens L, Yang-Feng TL, Liao YC, Chen E, Gray A, McGrath J, Seeburg PH, Libermann TA, Schlessinger J, Franke U, Levinson A, Ullrich A: Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene. Science 230:1132–1139, 1985
Bargmann CI, Hung MC, Weinberg RA: The neu oncogene encodes an epidermal growth factor receptor related protein. Nature 319:226–230, 1986
Stern DF, Heffernan PA, Weinberg RA: p185, a product of the neu-proto-oncogene, is a receptor like protein associated with tyrosine kinase. Mol Cell Biol 6:1728–1740, 1986
Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL: Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235:177–182, 1987
Slamon DJ, Clark GM: Amplification of c-erbB-2 and aggressive human breast tumor. Science 240:1795–1798, 1988
Kraus MH, Popescu NC, Amsbaug C, King CR: Over-expression of the EGF-receptor related proto-oncogene erbB-2 in human mammary tumor cell lines by different molecular mechanisms. EMBO J 6:605–610, 1987
Scatchard G: The attractions of proteins for small molecules and ions. Ann NY Acad Sci 51:660–672, 1949
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275, 1951
Sun TT, Green H: Keratin cytoskeleton in epithelial cells of internal organs. Proc Natl Acad Sci USA 76:2813–2817, 1979
Schlegel R, Banks-Schlegel S, Pinkus GS: Immunohistochemical localization of keratin in normal human tissues. Lab Invest 42:91–95, 1980
Fuchs E, Green H: The expression of keratin genes in epidermis and cultured epidermal cells. Cell 15: 887–897, 1978
Calaf G, Russo IH, Roi L, Russo J: Effect of estrogen on the length of S phase of human breast tissue in organ culture. IRCS Med Sci 10:307–308, 1982
Calaf G, Russo IH, Roi L, Russo J: Hormonal response of human breast tissue in organ culture. IRCS Med Sci 10:566–567, 1982
Calaf G, Russo IH, Roi L, Russo J: Effects of peptides and steroid hormones on cell kinetic parameters of normal human breast tissue in organ culture. In Vitro Cell Develop Biol 22:135–140, 1986
Borg A, Tandon AK, Sigurdsson H, Clark GM, Ferno M, Fuqua SAW, Killander O, McGuire WL: HER-2/neu amplification predicts poor survival in node positive breast cancer. Cancer Res 50:4332–4337, 1990
Yokota J, Yokota YT, Battifora H, Fevre CL, Cline MJ: Alterations of myc, myb and ras Ha protooncogenes in cancer are frequent and show clinical correlations. Science 231:261, 1986
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Calaf, G., Abarca-Quinones, J., Feuilhade, F. et al. New cell lines of human breast cancer origin. Breast Cancer Res Tr 21, 63–75 (1992). https://doi.org/10.1007/BF01811965
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DOI: https://doi.org/10.1007/BF01811965