This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Slovak, M. L. Articles by Cole, S. P.C. Search for Related Content PubMed PubMed Citation Articles by Slovak, M. L. Articles by Cole, S. P.C.

Localization of a Novel Multidrug Resistance-associated Gene in the HT1080/DR4 and H69AR Human Tumor Cell Lines¹

Department of Cytogenetics, City of Hope National Medical Center, Duarte, California 91010 [M. L. S., J. P. H.], and Cancer Research Laboratories, Queen's University, Kingston, Ontario, Canada K7L 3N6 [G. B., E. U. K., R. G. D., S. P. C. C.]

Two doxorubicin-selected human tumor cell lines, H69AR and HT1080/DR4, display a multidrug resistance phenotype but do not overexpress P-glycoprotein. Recently, a 6.5-kilobase mRNA encoding a novel member of the ATP-binding cassette superfamily of transport proteins, designated multidrug resistance-associated protein (MRP), has been identified in the H69AR cell line. In the present study, the levels of MRP mRNA were found to be 14-fold higher in HT1080/DR4 cells relative to sensitive HT1080 cells. Southern blotting indicates that gene amplification contributes to the overexpression of MRP in HT1080/DR4 cells. Using a 4-kilobase MRP complementary DNA probe, MRP genes were localized to 2–5 chromosomes bearing homogeneously staining regions and to multiple double minute chromosomes in H69AR cells. Resistant H69AR cells also contained a new der(16) with a structural aberration affecting 16p13.1, the normal cellular locus of the MRP gene. The MRP probe hybridized to two small homogeneously staining regions (hsr) in HT1080/DR4 cells including hsr(7)(p12p15). MRP localization was restricted to the normal cellular locus, 16p13.1, in the parental H69 and HT1080 cells and the drug-sensitive H69PR revertant cells. Our data provide combined evidence that amplification of the MRP gene is associated with the expression of drug resistance in selected solid tumor cell lines.

¹ This work was supported in part by Grants CA-33572 (M. L. S.), and the Medical Research Council (MRC) of Canada (R. G. D., S. P. C. C.). E. U. K. is the recipient of an MRC studentship.

² To whom requests for reprints should be addressed, at City of Hope National Medical Center, Department of Cytogenetics, Northwest Bldg., Room 2255, 1500 E. Duarte Road, Duarte, CA 91010.

Received 4/ 9/93. Accepted 6/14/93.

This article has been cited by other articles:

				M. Fryknas, L. Rickardson, M. Wickstrom, S. Dhar, H. Lovborg, J. Gullbo, P. Nygren, M. G. Gustafsson, A. Isaksson, and R. Larsson Phenotype-based screening of mechanistically annotated compounds in combination with gene expression and pathway analysis identifies candidate drug targets in a human squamous carcinoma cell model. J Biomol Screen, August 1, 2006; 11(5): 457 - 468. [Abstract] [PDF]

				R. G. Deeley, C. Westlake, and S. P. C. Cole Transmembrane Transport of Endo- and Xenobiotics by Mammalian ATP-Binding Cassette Multidrug Resistance Proteins. Physiol Rev, July 1, 2006; 86(3): 849 - 899. [Abstract] [Full Text] [PDF]

				M. Muredda, K.-I. Nunoya, R. A. Burtch-Wright, E. U. Kurz, S. P. C. Cole, and R. G. Deeley Cloning and Characterization of the Murine and Rat mrp1 Promoter Regions Mol. Pharmacol., November 1, 2003; 64(5): 1259 - 1269. [Abstract] [Full Text] [PDF]

				G. Bacher, B. Nickel, P. Emig, U. Vanhoefer, S. Seeber, A. Shandra, T. Klenner, and T. Beckers D-24851, a Novel Synthetic Microtubule Inhibitor, Exerts Curative Antitumoral Activity in Vivo, Shows Efficacy toward Multidrug-resistant Tumor Cells, and Lacks Neurotoxicity Cancer Res., January 1, 2001; 61(1): 392 - 399. [Abstract] [Full Text]

				K. Toth, M. M. Vaughan, G. Schwartz, J. S. Winston, B. S. Skenderis II, H. K. Slocum, and Y. M. Rustum Expression of the MRP and MDRI Multidrug Resistance Gene Products in 160 Untreated Human Carcinomas Studied by Immunohistochemical Methods in Formalin-Paraffin Sections International Journal of Surgical Pathology, July 1, 1998; 6(3): 145 - 154. [Abstract] [PDF]

				C. Millot, J.-M. Millot, H. Morjani, A. Desplaces, and M. Manfait Characterization of Acidic Vesicles in Multidrug-resistant and Sensitive Cancer Cells by Acridine Orange Staining and Confocal Microspectrofluorometry J. Histochem. Cytochem., September 1, 1997; 45(9): 1255 - 1264. [Abstract] [Full Text] [PDF]

				Z. Zhan, V. A. Sandor, E. Gamelin, J. Regis, B. Dickstein, W. Wilson, A. T. Fojo, and S. E. Bates Expression of the Multidrug Resistance-Associated Protein Gene in Refractory Lymphoma: Quantitation by a Validated Polymerase Chain Reaction Assay Blood, May 15, 1997; 89(10): 3795 - 3800. [Abstract] [Full Text] [PDF]

				D. W. Loe, K. C. Almquist, S. P. C. Cole, and R. G. Deeley ATP-dependent 17beta-Estradiol 17-(beta-D-Glucuronide) Transport by Multidrug Resistance Protein (MRP) J. Biol. Chem., April 19, 1996; 271(16): 9683 - 9689. [Abstract] [Full Text] [PDF]

				S. Ruetz, M. Brault, C. Kast, C. Hemenway, J. Heitman, C. E. Grant, S. P. C. Cole, R. G. Deeley, and P. Gros Functional Expression of the Multidrug Resistance-associated Protein in the Yeast Saccharomyces cerevisiae J. Biol. Chem., February 23, 1996; 271(8): 4154 - 4160. [Abstract] [Full Text] [PDF]

				M. D. Norris, S. B. Bordow, G. M. Marshall, P. S. Haber, S. L. Cohn, and M. Haber Expression of the Gene for Multidrug-Resistance-Associated Protein and Outcome in Patients with Neuroblastoma N. Engl. J. Med., January 25, 1996; 334(4): 231 - 238. [Abstract] [Full Text] [PDF]