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MDR1mRNA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MDR1mRNA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MDR1mRNA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。
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 ceils 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.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2013年第6期865-870,共6页
Journal of Sichuan University(Medical Sciences)
基金
四川省科技计划项目(No.2012SZ0022)
长江学者和创新团队发展计划(No.IRT0935)资助
