Down-regulation of Ras-related Protein Rab 5C-dependent Endocytosis and Glycolysis in Cisplatin-resistant Ovarian Cancer Cell Lines
Down-regulation of Ras-related Protein Rab 5C-dependent Endocytosis and Glycolysis in Cisplatin-resistant Ovarian Cancer Cell Lines
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Ras 相关蛋白 Rab 5C 依赖性内吞作用和糖酵解作用在顺铂耐药卵巢癌细胞系中的下调
DOI：
10.1074/mcp.m113.033217
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发表时间：
2014-11-01
影响因子：
7
通讯作者：
Deng, Haiteng
中科院分区：
文献类型：
--
作者：
Jin, Lixu;Huo, Yi;Deng, Haiteng
耐药性对卵巢癌的治疗提出了重大挑战。了解耐药机制对于寻找新的治疗靶点非常重要。本工作建立了顺铂耐药的卵巢癌细胞系A2780-DR，耐药指数为6.64。与A2780细胞相比，A2780-DR细胞中顺铂的细胞积累显着减少，这与耐药性的一般特征一致。定量蛋白质组学分析鉴定了 A2780 和 A2780-DR 细胞之间的 340 个差异表达蛋白，这些蛋白涉及多种细胞过程，包括代谢过程、细胞成分生物发生、细胞过程和应激反应。实时定量PCR和Western blotting证实A2780-DR细胞中Ras相关蛋白Rab 5C和Rab 11B的表达水平低于A2780细胞。 A2780 细胞中短发夹 (sh)RNA 介导的 Rab 5C 敲低导致对顺铂的耐药性显着增加，而 A2780-DR 细胞中 Rab 5C 的过表达增加了对顺铂的敏感性，表明 Rab 5C 依赖性内吞作用在顺铂耐药性中发挥着重要作用。我们的结果还表明，耐药细胞中糖酵解酶丙酮酸激酶、葡萄糖-6-磷酸异构酶、果糖二磷酸醛缩酶、乳酸脱氢酶和磷酸甘油酸激酶1的表达下调，表明卵巢癌的耐药性与糖酵解的减少直接相关。此外，还发现 A2780-DR 中谷胱甘肽还原酶上调，而波形蛋白、HSP90 以及膜联蛋白 A1 和 A2 下调。综上所述，我们的结果表明卵巢癌细胞系 A2780 的耐药性是由多因素引起的，包括 Rab 5C 依赖性顺铂、糖酵解酶和波形蛋白内吞作用的下调，以及抗氧化蛋白的上调，表明 Rab 5C 是治疗耐药性卵巢癌的潜在靶点。这为全面了解卵巢癌耐药性又迈出了一步。
Drug resistance poses a major challenge to ovarian cancer treatment. Understanding mechanisms of drug resistance is important for finding new therapeutic targets. In the present work, a cisplatin-resistant ovarian cancer cell line A2780-DR was established with a resistance index of 6.64. The cellular accumulation of cisplatin was significantly reduced in A2780-DR cells as compared with A2780 cells consistent with the general character of drug resistance. Quantitative proteomic analysis identified 340 differentially expressed proteins between A2780 and A2780-DR cells, which involve in diverse cellular processes, including metabolic process, cellular component biogenesis, cellular processes, and stress responses. Expression levels of Ras-related proteins Rab 5C and Rab 11B in A2780-DR cells were lower than those in A2780 cells as confirmed by real-time quantitative PCR and Western blotting. The short hairpin (sh)RNA-mediated knockdown of Rab 5C in A2780 cells resulted in markedly increased resistance to cisplatin whereas overexpression of Rab 5C in A2780-DR cells increases sensitivity to cisplatin, demonstrating that Rab 5C-dependent endocytosis plays an important role in cisplatin resistance. Our results also showed that expressions of glycolytic enzymes pyruvate kinase, glucose-6-phosphate isomerase, fructose-bisphosphate aldolase, lactate dehydrogenase, and phosphoglycerate kinase 1 were down-regulated in drug resistant cells, indicating drug resistance in ovarian cancer is directly associated with a decrease in glycolysis. Furthermore, it was found that glutathione reductase were up-regulated in A2780-DR, whereas vimentin, HSP90, and Annexin A1 and A2 were down-regulated. Taken together, our results suggest that drug resistance in ovarian cancer cell line A2780 is caused by multifactorial traits, including the down-regulation of Rab 5C-dependent endocytosis of cisplatin, glycolytic enzymes, and vimentin, and up-regulation of antioxidant proteins, suggesting Rab 5C is a potential target for treatment of drug-resistant ovarian cancer. This constitutes a further step toward a comprehensive understanding of drug resistance in ovarian cancer.