miR-130a对卵巢癌A2780细胞顺铂耐药性的影响及其机制的研究

Regulatory Effects and Associated Mechanisms of miR-130a Molecules on Cisplatin Resistance in Ovarian

中文关键词:  卵巢癌 顺铂耐药 miR-130a 抑癌基因（PTEN） 多耐药基因1(MDR1) P-糖蛋白(P-gp）

英文关键词:Ovarian cancer Cisplatin resistance miR-130a PTEN MDR1 P-gp

中文摘要:

      目的 探讨miR-130a表达的改变对卵巢癌A2780细胞（包括顺铂敏感细胞株A2780s和耐药株A2780/DDP）顺铂耐药性的影响及其机制。方法 将A2780s、A2780/DDP细胞各分为4组，分别予以单纯脂质体处理、转染阴性对照小RNA、miR-130a模拟物(可使miR-130a表达增加)、miR-130a抑制物(降低miR-130a表达)处理，MTS法检测各组细胞增殖情况和对顺铂的耐药性，RT-PCR、Western blot 法检测未处理和处理后细胞多耐药基因1（MDR1）、抑癌基因（PTEN） mRNA和蛋白的表达。结果 A2780/DDP细胞MDR1 mRNA和MDR1的表达产物P-糖蛋白(P-gp）的表达高于A2780s细胞（P<0.05），而PTEN mRNA及蛋白的表达在两者间差异无统计学意义。上调miR-130a的表达对A2780s和A2780/DDP细胞的增殖无直接影响，但能增强其对顺铂的耐药性，且提高MDR1 mRNA和P-gp表达水平；下调miR-130a的表达，同样不影响细胞的增殖, 但增强其对顺铂的敏感性, 并可降低MDR1 mRNA和P-gp的表达，提高PTEN蛋白的表达。结论 miR-130a抑制物通过上调PTEN蛋白和下调P-gp的表达来逆转卵巢癌A2780细胞系对顺铂的耐药性。miR-130a有望成为耐药性卵巢癌基因治疗的新靶点。

英文摘要:

      Objective To determine the regulatory effects and associated mechanisms of miR-130a on cisplatin resistance in ovarian cancer A2780 cell lines （including cisplatin sensitive A2780s and its resistant A2780/DDP cells）. Methods A2780s and A2780/DDP cells were divided into four groups, and treated with lipo2000 (Lip), miR-negative （miR-NC） control, miR-130a-mimics (miR-130a-M increasing the expression of miR-130a and the agent), and miR-130a-inhibitor (miR-130a-I downregulating miR-130a expression), respectively. The proliferation of cells and their sensitivity to cisplatin were detected by MTT assay. RT-PCR and western blot were performed to examine the levels of MDR1, PTEN mRNA and proteins. Results The expressions of MDR1 mRNA and P-gp in the A2780/DDP cells were significantly higher than those in the A2780s cells. However, no differences in the expressions of PTEN mRNA and proteins were detected between the two cell lines. Over-expressions of miR-130a had no effect on cell proliferation, but increased the resistance of the cells to cisplatin and up-regulated the expressions of MDR1 mRNA and P-gp in both cell lines. Down-regulated miR-130a did not affect cell proliferations, but enhanced the sensitivity of the cells to cisplatin, inhibited the expressions of MDR1 mRNA and P-gp and increased the expression of PTEN proteins. Conclusion MiR-130a expression may be associated with cisplatin resistance of ovarian cancer cells. MiR-130a inhibitor can reverse the cisplatin resistance by up-regulating the expression of PTEN proteins and down-regulating P-gp in A2780 cell lines. MiR-130 may become a new potential target of genetic therapy for cisplatin-resistant ovarian cancers.