Li Ning-wei, Wang Hong-Jing, Yang Ling-Yun, Jia Xi-Biao, Chen Cen, Wang Xue
Department of Obstetrics and Gynecology, West China Second Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2013 Nov;44(6):865-70.
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).
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.
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.
MiR-130a expression may be associated with cisplatin resistance of ovarian cancer cells. MiR-130a inhibitor can reverse the cisplatin resistance by upregulating 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.
确定miR-130a对卵巢癌A2780细胞系（包括顺铂敏感的A2780s及其耐药的A2780/DDP细胞）顺铂耐药性的调控作用及相关机制。
将A2780s和A2780/DDP细胞分为四组，分别用脂质体2000（Lip）、miR阴性（miR-NC）对照、miR-130a模拟物（miR-130a-M，增加miR-130a表达）和miR-130a抑制剂（miR-130a-I，下调miR-130a表达）处理。通过MTT法检测细胞增殖及其对顺铂的敏感性。进行RT-PCR和蛋白质印迹法检测MDR1、PTEN mRNA和蛋白质水平。
A2780/DDP细胞中MDR1 mRNA和P-gp的表达明显高于A2780s细胞。然而，两种细胞系之间PTEN mRNA和蛋白质的表达未检测到差异。miR-130a的过表达对细胞增殖无影响，但增加了细胞对顺铂的耐药性，并上调了两种细胞系中MDR1 mRNA和P-gp的表达。下调miR-130a不影响细胞增殖，但增强了细胞对顺铂的敏感性，抑制了MDR1 mRNA和P-gp的表达，并增加了PTEN蛋白质的表达。
miR-130a表达可能与卵巢癌细胞的顺铂耐药性有关。miR-130a抑制剂可通过上调A2780细胞系中PTEN蛋白质的表达和下调P-gp来逆转顺铂耐药性。miR-130可能成为顺铂耐药卵巢癌基因治疗的新潜在靶点。
