Abstract

The ability of tumor cells to develop simultaneous resistance to multiple cytotoxic drugs constitutes a major problem in cancer chemotherapy. This multidrug resistant phenotype is due to decreased intracellular drug accumulation, apparently as a result of alterations in plasma membrane. It was recently demonstrated by us that multidrug resistance in cultured Chinese hamster cells correlates with amplification of specific DNA sequences. An amplified gene associated with multidrug resistance in these cells was isolated. Recently DNA sequences homologous to this gene were found to be amplified in leukemic cells of a chemotherapy-resistant patient. In order to investigate the genetic mechanisms involved in the development of multidrug resistance by human tumors, the human homologue of the Chinese hamster gene will be isolated, and its amplification and expression in multidrug-resistant human tumor cells will be analyzed. In addition, multidrug-resistant human tumor cells will be assayed for amplification of other genes, using the in-gel DNA renaturation technique developed by the P.I. This technique detects amplified genes in the absence of any preliminary information about their sequence or location in the genome. Any additional amplified genes found by this procedure will be cloned and analyzed. Amplification and expression of the cloned genes in chemotherapy-resistant human tumor samples will be correlated with the tumor drug resistance profiles. Tumors that have acquired drug resistance in the course of chemotherapy and tumors initially unresponsive to chemotherapy will be compared with regard to the expression of cloned genes. Expression of these genes in individual cells within the tumors will be analyzed by in situ hybridization. Full-length cDNA and/or genomic clones of multidrug resistance-associated genes will be isolated, and the functions of these genes will be analyzed by gene transfer. cDNA sequences of the cloned genes will be determined. The products of these genes will be expressed in bacteria. At a later stage, these products or chemically synthesized peptides will be used to raise antibodies against multidrug-resistant human tumor cells, with the aim of investigating the possible diagnostic and therapeutic applications for such antibodies.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA040333-01
Application #
3180143
Study Section
Experimental Therapeutics Subcommittee 2 (ET)
Project Start
1985-08-01
Project End
1988-07-31
Budget Start
1985-08-01
Budget End
1986-07-31
Support Year
1
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
University of Illinois at Chicago
Department
Type
Overall Medical
DUNS #
121911077
City
Chicago
State
IL
Country
United States
Zip Code
60612

  Publications

Tang-Wai, D F; Kajiji, S; DiCapua, F et al. (1995) Human (MDR1) and mouse (mdr1, mdr3) P-glycoproteins can be distinguished by their respective drug resistance profiles and sensitivity to modulators. Biochemistry 34:32-9
Gudkov, A V; Kazarov, A R; Thimmapaya, R et al. (1994) Cloning mammalian genes by expression selection of genetic suppressor elements: association of kinesin with drug resistance and cell immortalization. Proc Natl Acad Sci U S A 91:3744-8
Gudkov, A V; Zelnick, C R; Kazarov, A R et al. (1993) Isolation of genetic suppressor elements, inducing resistance to topoisomerase II-interactive cytotoxic drugs, from human topoisomerase II cDNA. Proc Natl Acad Sci U S A 90:3231-5
Chaudhary, P M; Roninson, I B (1993) Induction of multidrug resistance in human cells by transient exposure to different chemotherapeutic drugs. J Natl Cancer Inst 85:632-9
Schinkel, A H; Arceci, R J; Smit, J J et al. (1993) Binding properties of monoclonal antibodies recognizing external epitopes of the human MDR1 P-glycoprotein. Int J Cancer 55:478-84
Levine, E A; Holzmayer, T A; Roninson, I B et al. (1993) MDR-1 expression in metastatic malignant melanoma. J Surg Res 54:621-4
Frommel, T O; Coon, J S; Tsuruo, T et al. (1993) Variable effects of sodium butyrate on the expression and function of the MDR1 (P-glycoprotein) gene in colon carcinoma cell lines. Int J Cancer 55:297-302
Mechetner, E B; Roninson, I B (1992) Efficient inhibition of P-glycoprotein-mediated multidrug resistance with a monoclonal antibody. Proc Natl Acad Sci U S A 89:5824-8
Holzmayer, T A; Hilsenbeck, S; Von Hoff, D D et al. (1992) Clinical correlates of MDR1 (P-glycoprotein) gene expression in ovarian and small-cell lung carcinomas. J Natl Cancer Inst 84:1486-91
Chaudhary, P M; Mechetner, E B; Roninson, I B (1992) Expression and activity of the multidrug resistance P-glycoprotein in human peripheral blood lymphocytes. Blood 80:2735-9

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