Abstract
Background: Kinetic resistance to cytotoxic drugs may account for the moderate responsiveness of breast cancer to chemotherapy. In the present study the in vitro effects of estradiol-mediated DNA stimulation on the cellular uptake of the DNA intercalating drug doxorubicin (DOX) were examined in MCF-7 human breast cancer cells. Methods: Using the fluorescent properties of the drug, the cellular uptake was investigated by high performance liquid chromatography (HPLC), and by flow cytometry. Results: The uptake of DOX (0.01–2 μg/ml) by MCF-7 cells in suspension, incubated for 1 and 6 h, showed a strong correlation between the incubation concentration of DOX and the cellular uptake of the drug as measured by HPLC and flow cytometry. Simultaneous exposure of MCF-7 cells, in monolayer culture, to DOX (0.04–0.2 μg/ml) and estradiol (1 nM) for 1–24 h showed no significant difference in uptake of the drug compared to control cultures exposed to DOX in the absence of estradiol. Neither was there a significant difference in uptake of DOX when MCF-7 cells were pretreated with estradiol (1 nM) for 16–24 h followed by a 0.5, 1, 6, and 21/23 h incubation with DOX (0.01–2 μg/ml). Pretreatment with estradiol did not affect the retention of DOX as measured 24 h after a 0.5 h incubation with DOX (2 μg/ml). Furthermore, fluorescence microscopy revealed no difference in the cellular DOX distribution pattern of estradiol-stimulated MCF-7 cultures compared to unstimulated cultures. Conclusion: From this study we can conclude that, for the human MCF-7 breast cancer cells in vitro, the uptake, retention, and cellular distribution of DOX are not influenced by estrogenic manipulation.
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References
Launchbury AP, Habboubi N: Epirubicin and doxorubicin: a comparison of their characteristics, therapeutic activity and toxicity. Cancer Treat Rev 19: 197–228, 1993
The French Epirubicin Study Group: A prospective randomized phase III trial comparing combination chemotherapy with cyclophosphamide, fluorouracil, and either doxorubicin or epirubicin. J Clin Oncol 6: 679–688, 1988
Italian Multicenter Breast Study with Epirubicin: Phase III randomized study of fluorouracil, cpirubicin, and cyclophosphamide vs fluorouracil, doxorubicin, and cyclophos phamide in advanced breast cancer: An Italian multicenter trial. J Clin Oncol 6: 976–982, 1988
Levine M, Bramwell V, Pritchard K, Shepherd L: CEF versus CMF in premenopausal women — Recent update. Sixth International Conference on Adjuvant Therapy of Primary Breast Cancer, St Gallen, Switzerland, February 25–28, 1998, abstract S35
Liu LF, Rowe TC, Yang L, Tewey KM, Chen GL: Cleavage of DNA by mammalian DNA topoisomerase II. J Biol Chem 258: 15365–15370, 1983
Sinha BK, Katki AG, Bastist G, Cowan KH, Myers CE: Differential formation of hydroxyl radicals by adriamycin in sensitive and resistant MCF-7 human breast cells: implication for the mechanisms of action. Biochemistry 26: 3776–3781, 1987
Chabner BA, Myers CE: Antitumor antibiotics. In: DeVita V, Hellman S, Rosenberg S (eds) Cancer, Principles and Practice of Oncology, 4th edition. JB Lippincott, Philadelphia, 1993, pp 376–380
Beck WT: The cell biology of multiple drug resistance. Biochem Pharmacol 36: 2879–2887, 1987
Endicott JA, Ling V: The biochemistry of P-glycoprotein mediated multidrug resistance. Annu Rev Biochem 58: 137–171, 1989
Lehner M: Reversal of multidrug resistance in breast cancer: many more open questions than answers. Ann Oncol 4: 11–13, 1993
Bontenbal M, Sieuwerts AM, Klijn JGM, Peters HA, Krijnen HLJM, Sonneveld P, Fockens JA: Effect of hormonal manipulation and doxorubicin administration on cell kinetics of human breast cancer cells. Br J Cancer 60: 688–692, 1989
Weichselbaum RR, Hellman S, Piro AJ, Nove JJ, Little JB: Proliferation kinetics of a human breast cancer line in vitro following treatment with 17β-estradiol and 1-β-D-arabinofuranosylcytosine. Cancer Res 38: 2339–2342, 1978
Clarke R, van der Berg HW, Kennedy DJ, Murphy RF: Estrogen receptor status and the response of human breast cancer cell lines to a combination of methotrexate and 17β-estradiol. Br J Cancer 51: 365–369, 1985
Hug V, Johnston D, Finders M, Hortobagyi G: Use of growth stimulatory hormones to improve the in vitro therapeutic index of doxorubicin for human breast tumors. Cancer Res 46: 147–152, 1986
Bontenbal M, Sieuwerts AM, Peters HA, Sonneveld P, Foekens JA, Klijn JGM: Manipulation of cell cycle kinetics: influence on the cytotoxicity of doxorubicin in human breast cancer cells. J Steroid Biochem Molec Biol 37: 1097–1101, 1990
Remvikos Y, Jouve M, Beuzeboc P, Viehl P, Magdelenat H, Pouillart P: Cell cycle modifications of breast cancers during neo-adjuvant chemotherapy: a flow cytometry study on fine needle aspirates. Eur J Cancer 13: 1843–1848, 1993
Daidone MG, Silvestrini R, Valentinis B, Ferrari L, Bartoli C: Changes in cell kinetics induced by primary chemotherapy in breast cancer. Int J Cancer 47: 380–383, 1991
Israel M, Pegg WJ, Wilkinson PM, Garnick M: Liquid chromatographic analysis of adriamycin and metabolites in biological fluids. J Liquid Chromatography 6: 795–809, 1985
Kokenberg E, Sonneveld P, Delwel R, Sizoo W, Hagenbeek A, Löwenberg B: In vivo uptake of daunorubicin by acute myeloid leukemia (AML) cells measured by flow cytometry. Leukemia 2: 511–517, 1988
Citro G, Cucco C, Verdina A, Zupi G: Reversal of adriamycin resistance by lonidamine in a human breast cancer cell line. Br J Cancer 64: 534–536, 1991
Taylor CW, Dalton WS, Parrish PR, Gleason MC, Bellamy WT, Thompson FH, Roe DJ, Trent JM: Different mechanisms of decreased drug accumulation in doxorubicin and mitoxantrone resistant variants of the MCF7 human breast cancer cell line. Br J Cancer 63: 923–929, 1991
Schuurhuis GJ, Broxterman HJ, Cervantes A, van Heijningen ThHM, de Lange JHM, Baak JPA, Pinedo HM, Lankelma J: Quantitative determination of factors contributing to doxorubicin resistance in multidrug-resistant cells. J Natl Cancer Inst 81: 1887–1892, 1989
Tokita N, Raju MR: Cell cycle dependent adriamycin uptake in Chinese hamster cells. Eur J Cancer Clin Oncol 21: 243–246, 1985
Minderman H, Linssen P, Wessels J, Haanen C: Cell cycle related uptake, retention and toxicity of idarubicin, daunorubicin and doxorubicin. Anticancer Res 13: 1161–1166, 1993
Ling Y, El-Naggar AK, Priebe W, Perez-Soler R: Cell cycle-dependent cytotoxicity, G2M phase arrest, and disruption of p34cdc2/cyclin B1 activity induced by doxorubicin in synchronized P388 cells. Mol Pharmacol 49: 832–841, 1996
Epstein RJ, Smith PJ, Watson JV, Bleehan NM: Characterisation of VP-16-induced DNA cleavage in oestrogen stimulated human breast cancer cells. Br J Cancer 57: 445–450, 1988
Epstein RJ, Smith PJ: Estrogen-induced potentiation of DNA damage and cytotoxicity in human breast cancer cells treated with toposiomerase II-interactive antitumor drugs. Cancer Res 48: 297–303, 1988
Epstein RJ, Smith PJ, Watson JV, Waters C, Bleehan NM: Oestrogen potentiates topoisomerase-II-mediated cytotoxicity in an activated subpopulation of human breast cancer cells: Implications for cytotoxic drug resistance in solid tumours. Int J Cancer 44: 501–505, 1989
Shaikh NA, Owen AM, Ghilchik MW, Braunsberg H: Actions of medroxyprogesterone acetate on the efficacy of cytotoxic drugs: Studies with human breast cancer cells in culture. Int J Cancer 43: 458–463, 1989
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
Reed MJ, Ross MS, Ghilchik MW: The effect of oestrogens and medroxyprogesterone acetate on the uptake of cytotoxic drugs by MCF-7 breast cancer cells. Anticancer Res 12: 533–536, 1992
Osborne CK, Kitten L, Arteaga CL: Antagonism of chemotherapy-induced cytotoxicity for human breast cancer cells by antiestrogens. J Clin Oncol 7: 710–717, 1989
Jancis EM, Carbone R, Loechner KJ, Dannies PS: Estradiol induction of rhodamine 123 efflux and the multidrug resistance pump in rat pituitary tumor cells. Mol Pharmacol 43: 51–56, 1992
Bontenbal M, Sonneveld P, Fockens JA, Klijn JGM: Oestradiol enhances doxorubicin uptake and cytotoxicity in human breast cancer cells (MCF-7). Eur J Cancer Clin Oncol 24: 1409–1414, 1988
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Bontenbal, M., Sieuwerts, A.M., Peters, H.A. et al. Uptake and distribution of doxorubicin in hormone-manipulated human breast cancer cells in vitro. Breast Cancer Res Treat 51, 139–148 (1998). https://doi.org/10.1023/A:1006026015448
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DOI: https://doi.org/10.1023/A:1006026015448