Blockage of drug resistance in vitro by disulfiram, a drug used to treat alcoholism

被引:92
作者
Loo, TW
Clarke, DM
机构
[1] Univ Toronto, Dept Med, MRC, Grp Membrane Biol, Toronto, ON M5S 1A8, Canada
[2] Univ Toronto, Dept Biochem, MRC, Grp Membrane Biol, Toronto, ON M5S 1A8, Canada
来源
JNCI-JOURNAL OF THE NATIONAL CANCER INSTITUTE | 2000年 / 92卷 / 11期
关键词
D O I
10.1093/jnci/92.11.898
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Background: P-glycoprotein (P-gp) pumps a wide range of cytotoxic drugs out of cells. Inhibiting maturation of P-gp would be a novel method for circumventing P-gp-mediated multidrug resistance, which complicates cancer chemotherapy and treatment of patients infected with human immunodeficiency virus. We examined the effect of disulfiram (Antabuse(TM)) on the maturation and activity of P-gp, Methods: Embryonic kidney cells were transfected with a complementary DNA for the P-pg gene, and the effects of disulfiram on the sensitivity of the transfected cells to cytotoxic agents were determined. Enzyme assays were used to determine the effects of disulfiram on the verapamil-stimulated adenosine triphosphatase (ATPase) activity of P-gp, Disulfiram modifies cysteine residues, and mutant forms of P-gp that lack individual cysteines were used to determine whether particular cysteine residues mediate disulfiram's effects on P-gp activity. Maturation of recombinant P-gp was followed on immnnoblots. Results: Disulfiram increased the sensitivity of P-gp-transfected cells to vinblastine and colchicine and inhibited P-gp's verapamil-stimulated ATPase activity. Half-maximal inhibition of ATPase activity occurred at 13.5 mu M disulfiram, Disulfiram (at 100 mu M) inhibited a P-gp mutant by 43% (95% confidence interval [CI] = 37% - 48%) when cysteine was present at position 431 only and by 72% (95% CI = 66% - 77%) when cysteine was present at position 1074 only. Treatment of P-gp-transfected cells with 50 nM disulfiram blocked maturation of recombinant P-gp, Conclusions: Disulfiram can potentially reduce P-gp-mediated drug resistance by inhibiting P-gp activity (possibly via cysteine modification) and/or by blocking its maturation. These results suggest that disulfiram has the potential to increase the efficacy of drug therapies for cancer and acquired immunodeficiency syndrome.
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收藏
页码:898 / 902
页数:5
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