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The effects of cyclosporin A, tamoxifen, and medroxyprogesterone acetate on the enhancement of Adriamycin cytotoxicity in primary cultures of human breast epithelial cells

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Summary

Adriamycin (Adr), the single most active agent used in the treatment of breast cancer, may become ineffective as treatment progresses due to the development of multidrug resistant (MDR) tumors. A major mechanism associated with MDR is increased P-glycoprotein (Pgp) expression. This study examined the abilities of the anti-estrogen tamoxifen (TAM) and the progestin medroxyprogesterone acetate (MPA) as well as cyclosporin A (CsA), a known resistance modifier, to enhance the cytotoxic effects of Adr on human breast epithelial cells (HBEC) in primary culture. Pgp and estrogen receptor (ER) expression were determined in each of the cultures by immunocytochemical assays using the monoclonal antibodies C219 and H222 Spγ, respectively. The Adr-sensitive, Pgp-, ER+ MCF-7 cell line and the Adr-resistant, Pgp+, ER-MCF7-AdrR cell line were used as controls. Primary cultures were categorized as HBEC from tissues with or without previous chemotherapy. Pgp was detected in 1 of the 15 cell cultures from tissues without previous chemotherapy and in 5 of the 6 cell cultures from tissues previously exposed to chemotherapy. Incubation with either CsA or MPA plus Adr enhanced Adr toxicity in Pgp+ but not Pgp- cell cultures, whereas TAM had no effect on the sensitivity of any of the cultures. Of the 21 primary cultures of HBEC, 3 were ER+. There was no correlation between the enhancement of Adr cytotoxicity and ER status. The data suggest that MPA as well as CsA may be useful as modifying agents in overcoming Pgp-associated multidrug resistance.

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References

  1. Tormey DC: Adriamycin (NSC-123127) in breast cancer: an overview of studies. Cancer Chemother Rep 6: 319–327, 1975

    Google Scholar 

  2. Gottesman MM: How cancer cells evade chemotherapy. Cancer Res 53: 747–754, 1993

    PubMed  Google Scholar 

  3. Dalton WS: Mechanisms of drug resistance in breast cancer. Sem Oncol 14: 37–39, 1990

    Google Scholar 

  4. Whelan RDH, Waring CJ, Wolf CR, Hayes JD, Hosking LK, Hill BT: Over-expression of P-glycoprotein and glutathione S-transferase pi in MCF-7 cells selected for vincristine resistancein vitro. Int J Cancer 52: 241–246, 1992

    PubMed  Google Scholar 

  5. Veneroni S, Zaffaroni N, Daidone MG, Benini E, Villa R, Silvestrini R: Expression of p-glycoprotein andin vitro or in vivo resistance to doxorubicin and cisplatin in breast and ovarian cancers. Eur J Cancer 30A: 1002–1007, 1994

    PubMed  Google Scholar 

  6. Van der Valk P, van Kalken CK, Ketelaars H, Broxterman HJ, Scheffer G, Kuiper CM, Tsuruo T, Lankelma J, Meijer CJLM, Pinedo HM, Scheper RJ: Distribution of multidrug resistance-associated p-glycoprotein in normal and neoplastic human tissues. Annals of Oncol 1: 56–64, 1990

    Google Scholar 

  7. Schneider SL, Fuqua SAW, Speeg KV, Tandon AK, McGuire WL: Isolation and characterization of an adriamycin-resistant breast tumor cell line.In Vitro Cell Dev Biol 26: 621–628, 1990

    PubMed  Google Scholar 

  8. Leonessa F, Jacobson M, Boyle B, Lippman J, McGarvey M, Clarke R: Effect of tamoxifen on the multidrug-resistant phenotype in human breast cancer cells: isobologram, drug accumulation, and MT 170,000 glycoprotein (gp 170) binding studies. Cancer Res 54: 441–447, 1994

    PubMed  Google Scholar 

  9. Ries F, Dicato M: Treatment of advanced and refractory breast cancer with doxorubicin, vincristine and continuous infusion of verapamil. A phase I-II clinical trial. Med Oncol Tumor Pharmacother 8: 39–43, 1991

    PubMed  Google Scholar 

  10. Keilhauer G, Emling F, Raschack M, Gries J, Schlick E: The use of R-verapamil (R-VPM) is superior to racemic VPM in breaking multidrug resistance (MDR) of malignant cells (Abstract). Proc Amer Assoc Cancer Res 30: 2002, 1989

    Google Scholar 

  11. Erlichman C, Moore M, Thiessen JJ, Kerr IG, Walker S, Goodman P, Bjarnason G, DeAngelis C, Bunting P: Phase I pharmacokinetic study of cyclosporin A combined with doxorubicin. Cancer Res 53: 4837–4842, 1993

    PubMed  Google Scholar 

  12. Naito M, Yusa K, Tsuruo T: Steroid hormones inhibit binding of vinca alkaloid to multidrug resistance related p-glycoprotein. Biochem Biophys Res Comm 158: 1066–1071, 1989

    PubMed  Google Scholar 

  13. Yang CPH, DePinho SG, Greenberger LM, Arceci RJ, Horwitz SB: Progesterone interacts with p-glycoprotein in multidrug-resistant cells and in the endometrium of gravid uterus. J Biol Chem 264: 782–788, 1989

    PubMed  Google Scholar 

  14. Fine RL, Sachs CW, Albers ME, Safa A, Rao US, Scarborough G, Burchette J, Jordan C, Trump DL: Inhibition of multidrug resistance in human cancer cells by tamoxifen: laboratory to clinical studies. In: Miyazaki T, Takaku F, Sakuranda K (eds) The mechanism and new approach on drug resistance of cancer cells. Excerpta Medica, Amsterdam, 1993, pp 323–332

    Google Scholar 

  15. Ishida Y, Shimada Y, Shimoyama M: Synergistic effect of cyclosporin A and verapamil in overcoming vincristine resistance of multi-drug resistant cultured human leukemia cells. Jpn J Cancer Res 81: 834–843, 1990

    PubMed  Google Scholar 

  16. Tamai I, Safa AR: Competitive interaction of cyclosporins with the vinca alkaloid-binding site of p-glycoprotein in multidrug-resistant cells. J Biol Chem 265: 16509–16513, 1990

    PubMed  Google Scholar 

  17. Twentyman PR, Wright KA: Chemosensitisation of a drugsensitive parental cell line by low-dose cyclosporin A. Cancer Chemother Pharmacol 29: 24–28, 1991

    PubMed  Google Scholar 

  18. Emerman JT, Fiedler EE, Tolcher AW, Rebbeck PM: Effect of defined medium, fetal bovine serum, and human serum on growth and chemosensitivities of human breast cancer cells in primary culture: inference forin vitro assays. In Vitro Cell Dev Biol 23: 134–140, 1987

    PubMed  Google Scholar 

  19. Chan HS, Bradley G, Thorner P, Haddad G, Gallie BL, Ling V: A sensitive method for immunocytochemical detection of p-glycoprotein in multidrug-resistant human ovarian carcinoma cell lines. Lab Invest 59: 870–875, 1988

    PubMed  Google Scholar 

  20. Emerman JT, Tolcher AW, Rebbeck PM:In vitro sensitivity testing of human breast cancer cells to hormones and chemotherapeutic agents. Cancer Chemother Pharmacol 26: 245–249, 1990

    PubMed  Google Scholar 

  21. Emerman JT, Eaves CJ: Lack of effect of hematopoietic growth factors on human breast epithelial cell growth in serum-free primary culture. Bone Marrow Transplant 13: 285–291, 1994

    PubMed  Google Scholar 

  22. Emerman JT, Wilkinson DA: Routine culturing of normal, dysplastic and malignant human mammary epithelial cells from small tissue samples.In Vitro Cell Dev Biol 26: 1186–1194, 1990

    PubMed  Google Scholar 

  23. Gabelman BM, Emerman JT: Effects of estrogen, epidermal growth factor, and transforming growth factor-α on the growth of human breast epithelial cells in primary culture. Exp Cell Res 201: 113–118, 1992

    PubMed  Google Scholar 

  24. Schneider J, Bak M, Efferth Th, Kaufmann M, Mattern J, Volm M: P-glycoprotein expression in treated and untreated human breast cancer. Br J Cancer 60: 815–818, 1989

    PubMed  Google Scholar 

  25. Gundersen S, Kvinnsland S, Lundgren S, Klepp O, Lund E, Bormer O, Host H: Cyclical use of tamoxifen and high-dose medroxyprogesterone acetate in advanced estrogen receptor positive breast cancer. Breast Cancer Res Treat 17: 45–50, 1990

    PubMed  Google Scholar 

  26. Sedlacek SM, Horowitz KB: The role of progestins and progesterone receptors in the treatment of breast cancer. Steroids 44: 467–484, 1984

    PubMed  Google Scholar 

  27. Bojar H, Stuschke M, Staib W: Effects of high-dose medroxyprogesterone acetate on plasma membrane lipid mobility. Prog Cancer Res Ther 31: 115–119, 1984

    Google Scholar 

  28. Ramu A, Ramu N, Rosario LM: Circumvention of multidrug-resistance in P388 cells is associated with a rise in the cellular content of phosphatidylcholine. Biochem Pharmacol 41: 1455–1461, 1991

    PubMed  Google Scholar 

  29. O'Brian CA, Liskamp RM, Soloman DH, Weinstein IB: Triphenylethylenes: a new class of protein kinase Cinhibitors. J Natl Cancer Inst 76: 1243–1246, 1986

    PubMed  Google Scholar 

  30. Trump DL, Smith DC, Ellis PG, Rogers MP, Schold SC, Winer EP, Panella TJ, Jordan VC, Fine RL: High-dose oral Tamoxifen, a potential multidrug-resistance-reversal agent: Phase I trial in combination with vinblastine. J Natl Cancer Inst 84: 1811–1816, 1992

    PubMed  Google Scholar 

  31. Salmon SE, Grogan TM, Miller T, Scheper R, Dalton WS: Prediction of doxorubicin resistancein vitro in myeloma, lymphoma, and breast cancer by p-glycoprotein staining. J Natl Cancer Inst 81: 696–701, 1989

    PubMed  Google Scholar 

  32. Bergstraesser LM, Weitzman SA: Culture of normal and malignant primary human mammary epithelial cells in a physiological manner simulatesin vivo growth patterns and allows discrimination of cell type. Cancer Res 53: 2644–2654, 1993

    PubMed  Google Scholar 

  33. Petersen OW, Ronnov-Jessen L, Howlett AR, Bissell MJ: Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells. Proc Natl Acad Sci USA 89: 9064–9068, 1992

    PubMed  Google Scholar 

  34. Smith HS, Lippman ME, Hiller AJ, Stampfer MR, Hackett AJ: Response to doxorubicin of cultured normal and cancerous human mammary epithelial cells. J Natl Cancer Inst 74: 341–347, 1985

    PubMed  Google Scholar 

  35. Merkel DE, Fuqua SAW, Hill SM, McGuire WL: P-glycoprotein gene amplification or overexpression is not detected in clinical breast cancer specimens. Prog Clin Biol Res 276: 61–73, 1988

    PubMed  Google Scholar 

  36. Coradini D, Biffi A, Cappelletti V, Di Fronzo G: Activity of tamoxifen and new antiestrogens on estrogen receptor positive and negative breast cancer cells. Anticancer Res 14: 1059–1064, 1994

    PubMed  Google Scholar 

  37. Classen S, Possinger K, Pelka-Fleischer R, Wilmanns W: Effect of onapristone and medroxyprogesterone acetate on the proliferation and hormone receptor concentration of human breast cancer cells. J Steroid Biochem Mol Biol 45: 315–319, 1993

    PubMed  Google Scholar 

  38. Legha SS: Tamoxifen in the treatment of breast cancer. Ann Intern Med 109: 219–228, 1988

    PubMed  Google Scholar 

  39. Thomas M, Monet JD: Combined effects of RU486 and tamoxifen on the growth and cell cycle phases of the MCF-7 cell line. J Clin Endocrinol Metab 75: 865–870, 1992

    PubMed  Google Scholar 

  40. DeVleeschouwer N, Body JJ, Legros N, Muquardt C, Donnay I, Wouters P, Ledercq G: Growth factor-like activity of phenol red preparations in the MCF-7 breast cancer cell line. Anticancer Res 12: 789–794, 1992

    PubMed  Google Scholar 

  41. Chambers TC, Kuo JF: Phosphorylation and regulation of p-glycoprotein by protein kinase C. In: Miyazaki T, Takaku F, Sakuranda K (eds) The mechanism and new approach on drug resistance of cancer cells. Excerpta Medica, Amsterdam, 1993, pp 159–165

    Google Scholar 

  42. Kirk J, Houlbrook S, Stuart NSA, Stratford IJ, Harris AL, Carmichael J: Differential modulation of doxorubicin toxicity to multidrug and intrinsically drug resistant cell lines by anti-oestrogens and their major metabolites. Br J Cancer 67: 1189–1195, 1995

    Google Scholar 

  43. Shaikh NA, Owen AM, Ghilchik MW, Braunsberg H: Adriamycin action on human breast cancer cells: enhancement by medroxyprogesterone acetate. Int J Cancer 43: 733–736, 1989

    PubMed  Google Scholar 

  44. Kacinski BM, Yee LD, Carter D, Li D, Kuo MT: Human breast carcinoma levels of MDR-1 (P-glycoprotein) transcripts correlatein vivo inversely and reciprocally with tumor progesterone receptor content. Cancer Comm 1: 1–6, 1989

    Google Scholar 

  45. McGuire WL: P-glycoprotein and progesterone receptor in human breast cancer. Cancer Cells 2: 27–29, 1990

    PubMed  Google Scholar 

  46. Ishida H, Okabe M, Gomi K, Horiuchi R, Mikami K, Naito M, Tsuruo T: Modulation of adriamycin resistance in human breast carcinoma mcf-7 cellsin vitro andin vivo by medroxyprogesterone acetate. Jpn J Cancer Res 85: 542–549, 1994

    PubMed  Google Scholar 

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  1. Department of Anatomy, University of British Columbia, 2177 Wesbrook Mall, V6T 1Z3, Vancouver, B.C., Canada

    Jerome A. Claudio & Joanne T. Emerman

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  1. Jerome A. Claudio
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Claudio, J.A., Emerman, J.T. The effects of cyclosporin A, tamoxifen, and medroxyprogesterone acetate on the enhancement of Adriamycin cytotoxicity in primary cultures of human breast epithelial cells. Breast Cancer Res Tr 41, 111–122 (1996). https://doi.org/10.1007/BF01807156

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