INVOLVEMENT OF VACUOLAR H+-ADENOSINE TRIPHOSPHATASE-ACTIVITY IN MULTIDRUG RESISTANCE IN HL-60 CELLS
INVOLVEMENT OF VACUOLAR H+-ADENOSINE TRIPHOSPHATASE-ACTIVITY IN MULTIDRUG RESISTANCE IN HL-60 CELLS
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DOI：
10.1093/jnci/83.15.1098
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发表时间：
1991-08-07
影响因子：
10.3
通讯作者：
CENTER, MS
中科院分区：
文献类型：
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作者：
MARQUARDT, D;CENTER, MS
对长春新碱耐药的HL60细胞(HL60/VINC细胞)或阿霉素耐药的HL60细胞(HL60/ADR细胞)含有能量依赖的药物外排泵水平增强。HL60/VINC细胞含有药物转运蛋白P-糖蛋白，而HL60/ADR分离株则不含。在本研究中，我们研究了空泡H+-三磷酸腺苷酶(H+-ATPase)活性在HL60细胞耐药中的可能参与。我们使用了巴菲霉素A1，一种在低浓度下选择性抑制液泡H+-ATPase活性的药物。结果表明，巴菲霉素A1能显著增加HL60/ADR细胞和HL60/Venc细胞的药物蓄积，抑制药物外排。7-氯-4-硝基-2-氧杂-1，3-二氮唑也能抑制液泡H+-ATPase，也得到了类似的结果。F1F(O)线粒体ATPase抑制剂叠氮，以及E1E2型ATPase抑制剂钒酸和哇巴因对耐药细胞的药物水平没有影响。我们还观察到巴非霉素A1不与[H-3]叠氮多宾竞争结合P-糖蛋白。因此，巴菲霉素A1似乎不能作为P-糖蛋白的底物。提示空泡H+-ATPase活性参与了HL60/ADR细胞和HL60/Venc细胞的药物外排途径。这一作用的机制尚待确定。
HL60 cells isolated for resistance to vincristine (HL60/Vinc cells) or doxorubicin (HL60/Adr cells) contain enhanced levels of an energy-dependent drug efflux pump. HL60/Vinc cells contain the drug transporter P-glycoprotein, whereas the HL60/Adr isolate does not. In the present study, we examined the possible involvement of vacuolar H+-adenosine triphosphatase (H+-ATPase) activity in drug resistance in HL60 cells. We utilized bafilomycin A1, an agent which selectively inhibits vacuolar H+-ATPase activity at low concentrations. The results showed that bafilomycin A1 induced a major increase in drug accumulation and inhibited drug efflux in both HL60/Adr cells and HL60/Vinc cells. Similar results were obtained with 7-chloro-4-nitrobenz-2-oxa 1,3 diazole, an agent which is also capable of inhibiting vacuolar H+-ATPase. Azide, an inhibitor of F1F(o) mitochondrial ATPase, and vanadate and ouabain, which are inhibitors of E1E2-type ATPase, did not affect drug levels in resistant cells. We also observed that bafilomycin A1 did not compete with [H-3]azidopine binding to P-glycoprotein. Thus, bafilomycin A1 does not appear to function as a substrate for P-glycoprotein. These results suggest an involvement of vacuolar H+-ATPase activity in the pathway of drug efflux from HL60/Adr cells and HL60/Vinc cells. The mechanism of this action remains to be determined.