GS-X pump is functionally overexpressed in cis-diamminedichloroplatinum (II)-resistant human leukemia HL-60 cells and down-regulated by cell differentiation.
GS-X pump is functionally overexpressed in cis-diamminedichloroplatinum (II)-resistant human leukemia HL-60 cells and down-regulated by cell differentiation.
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DOI：
10.1016/s0021-9258(19)62016-8
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发表时间：
1994-11
期刊：
影响因子：
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通讯作者：
Toshihisa IshikawaS;Christine;Wright;H. Ishizuka
中科院分区：
文献类型：
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作者：
Toshihisa IshikawaS;Christine;Wright;H. Ishizuka
ATP 依赖性谷胱甘肽 S 结合物输出泵（称为 GS-X 泵）已被证明可以消除肿瘤细胞中潜在的细胞毒性谷胱甘肽-铂 (GS.Pt) 复合物，从而调节谷胱甘肽 (GSH) 相关的顺式二氯二氨铂 (II)（顺铂）耐药性（Ishikawa, T. 和 Ali-Osman, F. (1993) J.生物。268，20116-20125）。本研究提供的证据表明，GS-X 泵在顺铂耐药的人早幼粒细胞白血病 HL-60 (HL-60/R-CP) 细胞中功能性过度表达，其中细胞 GSH 水平显着增强。事实上，用质膜囊泡测量的 GS.Pt 复合物的 ATP 依赖性转运速率在 HL-60/R-CP 细胞中比在 HL-60 细胞中高约 4 倍。表观分子量分别为 200、110 和 70 kDa 的三种膜蛋白在 HL-60/R-CP 细胞中过表达，而在耐药细胞和敏感细胞的膜制剂中未通过免疫学检测到 P-糖蛋白 (MDR1)。与 HL-60 细胞不同，在 HL-60/R-CP 细胞的细胞质中观察到细胞内囊泡数量增加。使用 syn-(CICH2,CH3)-1,5-diazabicyclo-[3.3.0]-octa-3,6-dione-2,8-dione（单氯代二酮）的荧光显微镜显示，荧光谷胱甘肽 S-缀合物以能量依赖性方式积累在顺铂耐药细胞的细胞内囊泡中。 GS-X 泵被认为有助于囊泡介导的 GSH-药物缀合物从细胞中的排泄。此外，HL-60 和 HL-60/R-CP 细胞均因 12-O-十四烷酰佛波醇-13-乙酸酯、视黄酸和二甲亚砜而发生细胞分化，导致增殖停滞以及 c-myc mRNA 水平显着下降。细胞分化后，在由 HL-60/R-CP 和 HL-60 细胞制备的膜囊泡中，观察到 GS.Pt 复合物的 ATP 依赖性转运活性显着降低。这些结果表明GS-X泵在顺铂耐药和敏感细胞中的表达与细胞增殖相关。
The ATP-dependent glutathione S-conjugate export pump, named GS-X pump, has been shown to eliminate a potentially cytotoxic glutathione-platinum (GS.Pt) complex from tumor cells, thereby modulating glutathione (GSH)-associated resistance to cis-diamminedichloroplatinum(II) (cisplatin) (Ishikawa, T., and Ali-Osman, F. (1993) J. Biol. Chem. 268, 20116-20125). The present study provides evidence that the GS-X pump is functionally overexpressed in cisplatin-resistant human promyelocytic leukemia HL-60 (HL-60/R-CP) cells, in which the cellular GSH level was substantially enhanced. Indeed, the rate of ATP-dependent transport of the GS.Pt complex, measured with plasma membrane vesicles, was about 4-fold greater in HL-60/R-CP cells than in HL-60 cells. Three membrane proteins with apparent molecular masses of 200, 110, and 70 kDa were overexpressed in HL-60/R-CP cells, whereas P-glycoprotein (MDR1) was not immunologically detected in the membrane preparations from resistant and sensitive cells. Unlike in HL-60 cells, increased numbers of intracellular vesicles were observed in the cytoplasm of HL-60/R-CP cells. Fluorescence microscopy with syn-(CICH2,CH3)-1,5-diazabicyclo-[3.3.0]-octa-3,6-dione-2,8-dione (monochlorobimane) revealed that the fluorescent glutathione S-conjugate accumulated in intracellular vesicles of the cisplatin-resistant cells in an energy-dependent manner. The GS-X pump is suggested to contribute to vesicle-mediated excretion of GSH-drug conjugates from cells. In addition, both HL-60 and HL-60/R-CP cells underwent cell differentiation in response to 12-O-tetradecanoylphorbol-13-acetate, retinoic acid, and dimethyl sulfoxide, resulting in proliferation arrest as well as a remarkable decrease in the c-myc mRNA levels. After cell differentiation, a significant decrease was observed in the activity of ATP-dependent transport of the GS.Pt complex in membrane vesicles prepared from both HL-60/R-CP and HL-60 cells. These results suggest that the expression of the GS-X pump in both cisplatin-resistant and -sensitive cells is related to cell proliferation.