miR-485-5p靶向EGFR对卵巢癌顺铂耐药细胞系的影响

Effects of miR-485-5p on cisplatin-resistant ovarian cancer cell lines via targeting EGFR

目的探讨miR-485-5p对顺铂耐药卵巢癌细胞的影响及其机制。方法RT-qPCR检测人正常卵巢上皮细胞株(IOSE-80)及卵巢癌细胞株(A2780、SKOV3、OVCAR3、OVCA433)中miR-485-5p的表达。构建顺铂(DDP)耐药卵巢癌细胞并检测miR-485-5p与EGFR的表达。CCK8法检测各组细胞增殖能力,流式细胞术测定细胞凋亡。结果相对于IOSE-80细胞,各卵巢癌细胞株中miR-485-5p的表达显著下降,EGFR表达上升(均P<0.05)。SKOV3/DDP细胞株(0.17±0.02)中miR-485-5p表达较SKOV3细胞(0.32±0.04)进一步下降(t=5.81,P=0.004)。而SKOV3/DDP细胞中EGFR表达则较SKVO3进一步上升(P<0.05)。SKOV3/DDP细胞转染miR-485-5p mimic能抑制细胞增殖活力、诱导细胞凋亡,转染miR-485-5p inhibitor则相反(均P<0.05)。SKOV3/DDP细胞转染EGFR能促进细胞增殖并抑制细胞凋亡,该作用被miR-485-5p mimic部分抵消。结论miR-485-5p参与调控卵巢癌细胞的顺铂耐药,过表达miR-485-5p能提高卵巢癌化疗敏感性,该作用可能是通过负调控EGFR实现的。

Objective To explore the effects of miR-485-5p on cisplatin-resistant ovarian cancer cells and it’s mechanism.Methods RT-qPCR was used to detect the expression of miR-485-5p in human normal ovarian epithelial cell line(IOSE-80)and ovarian cancer cell lines(A2780,SKOV3,OVCAR3,OVCA433).Cisplatin(DDP)-resistant ovarian cancer cells were constructed and the expression of miR-485-5p and EGFR was detected.CCK8 assay was used to detect cell proliferation ability in each group.Cell apoptosis was measured by flow cytometry.Results Compared with IOSE-80 cell,miR-485-5p expression was decreased in each ovarian cancer cell lines,while the expression of EGFR was increased(all P<0.05).The expression of miR-485-5p in SKOV3/DDP cell lines was further decreased while EGFR expression was further increased than that in SKOV3 cells(P<0.05).Transfection of miR-485-5p mimic into SKOV3/DDP cells could inhibit cell proliferation and induce apoptosis,but the results were reversed when cells were transfected with miR-485-5p inhibitor(all P<0.05).The proliferation was increased while apoptosis was decreased in EGFR transfected SKOV3/DDP cells,but this effects was partially offseted by miR-485-5pmimic.Conclusion miR-485-5p participates in the regulation of cisplatin resistance of ovarian cancer cells,and overexpression of miR-485-5p can promote the chemosensitivity of ovarian cancer,which may be achieved through negative regulation of EGFR.