© 1999 by Oxford University Press
Journal of the National Cancer Institute, Vol. 91, No. 5, 429-433,
March 3, 1999
© 1999 Oxford University Press
ACCELERATED DISCOVERY |
Atypical Multidrug Resistance: Breast Cancer Resistance Protein Messenger RNA Expression in Mitoxantrone-Selected Cell Lines
Affiliations of authors: D. D. Ross, University of Maryland Greenebaum Cancer Center, Baltimore, Department of Medicine, Division of Hematology/Oncology, University of Maryland School of Medicine, and Baltimore Veterans Medical Center, Department of Veterans Affairs; W. Yang, University of Maryland Greenebaum Cancer Center; L. V. Abruzzo, University of Maryland Greenebaum Cancer Center and Department of Pathology, University of Maryland School of Medicine; W. S. Dalton, Moffitt Cancer Center, University of South Florida, Tampa; E. Schneider, Wadsworth Center, New York State Department of Health, Albany; H. Lage, M. Dietel, Institute of Pathology, University Hospital Charité, Humboldt University, Berlin, Germany; L. Greenberger, Wyeth-Ayerst Research, Pearl River, NY; S. P. C. Cole, Cancer Research Laboratories, Queen's University, Kingston, ON, Canada; L. A. Doyle, University of Maryland Greenebaum Cancer Center and Department of Medicine, Division of Hematology/Oncology, University of Maryland School of Medicine.
Correspondence to: Douglas D. Ross, M.D., Ph.D., Greenebaum Cancer Center of the University of Maryland, Rm. 9-015, Bressler Research Bldg., 655 West Baltimore St., Baltimore, MD 21201 (e-mail: DROSS{at}umcc01.umcc.ab. umd.edu). Reprint requests to: Douglas D. Ross or L. Austin Doyle, Greenebaum Cancer Center of the University of Maryland, Rm. 9-015, Bressler Research Bldg., 655 West Baltimore St., Baltimore, MD 21201.
BACKGROUND: Human cancer cell lines grown in the presence of the cytotoxic agent mitoxantrone frequently develop resistance associated with a reduction in intracellular drug accumulation without increased expression of the known drug resistance transporters P-glycoprotein and multidrug resistance protein (also known as multidrug resistance-associated protein). Breast cancer resistance protein (BCRP) is a recently described adenosine triphosphate-binding cassette transporter associated with resistance to mitoxantrone and anthracyclines. This study was undertaken to test the prevalence of BCRP overexpression in cell lines selected for growth in the presence of mitoxantrone. METHODS: Total cellular RNA or poly A+ RNA and genomic DNA were isolated from parental and drug-selected cell lines. Expression of BCRP messenger RNA (mRNA) and amplification of the BCRP gene were analyzed by northern and Southern blot hybridization, respectively. RESULTS: A variety of drug-resistant human cancer cell lines derived by selection with mitoxantrone markedly overexpressed BCRP mRNA; these cell lines included sublines of human breast carcinoma (MCF-7), colon carcinoma(S1 and HT29), gastric carcinoma (EPG85-257), fibrosarcoma (EPF86-079),and myeloma (8226) origins. Analysis of genomic DNA from BCRP-overexpressing MCF-7/MX cells demonstrated that the BCRP gene was also amplified in these cells. CONCLUSIONS: Overexpression of BCRP mRNA is frequently observed in multidrug-resistant cell lines selected with mitoxantrone, suggesting that BCRP is likely to be a major cellular defense mechanism elicited in response to exposure to this drug. It is likely that BCRP is the putative "mitoxantrone transporter" hypothesized to be present in these cell lines.
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