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目的:探讨miR-101表达对上皮性卵巢癌耐药细胞SKOV3/DDP行为活性的影响及相关作用机制。方法:上调SKOV3/DDP细胞中miR-101表达水平。RT-PCR法检测SKOV3/DDP细胞中miR-101 mRNA表达水平,CCK-8检测SKOV3/DDP细胞的顺铂敏感性,划痕实验检测SKOV3/DDP细胞的迁移力,Transwell实验检测SKOV3/DDP细胞侵袭力,Annexin V-FITC/PI流式细胞实验检测SKOV3/DDP细胞凋亡率,Western blot法检测SKOV3/DDP细胞p-ERK1/2、波形蛋白(Vimentin)、钙粘蛋白E(E-cadherin)及Caspase-3蛋白表达水平。结果:与各对照组比较,miR-101组SKOV3/DDP细胞miR-101mRNA表达水平明显升高(P<0.05),顺铂敏感性明显增加,IC50显著降低(P<0.05);划痕距离明显增宽(P<0.05),透膜细胞数显著减少(P<0.05);细胞凋亡率显著增高(P<0.05),p-ERK1/2及Vimentin蛋白表达水平降低(P<0.05),E-cadherin及Caspase-3表达水平显著升高(P<0.05)。结论:高表达的miR-101可抑制SKOV3/DDP细胞中p-ERK1/2蛋白表达,同时抑制细胞行为活性,促进顺铂诱导的细胞凋亡。
Objective: To investigate the effect of miR-101 on the behavioral activity of SKOV3 / DDP cells and its mechanism of action in epithelial ovarian cancer cells. Methods: Up-regulate the expression of miR-101 in SKOV3 / DDP cells. The expression of miR-101 mRNA in SKOV3 / DDP cells was detected by RT-PCR, the sensitivity of SKOV3 / DDP cells was detected by CCK-8, and the migration of SKOV3 / DDP cells was detected by scratch assay. The apoptosis rate of SKOV3 / DDP cells was detected by Annexin V-FITC / PI flow cytometry. The expressions of p-ERK1 / 2, vimentin and E-cadherin in SKOV3 / ) And Caspase-3 protein expression levels. Results: Compared with the control group, the miR-101mRNA expression level in SKOV3 / DDP cells was significantly increased (P <0.05), the sensitivity of cisplatin was significantly increased, and the IC50 was significantly decreased (P <0.05) (P <0.05), the number of transmembrane cells was significantly decreased (P <0.05), the apoptosis rate was significantly increased (P <0.05), the expression of p-ERK1 / 2 and Vimentin protein decreased -cadherin and Caspase-3 expression levels were significantly increased (P <0.05). CONCLUSION: High expression of miR-101 can inhibit the expression of p-ERK1 / 2 in SKOV3 / DDP cells, inhibit the activity of cells and promote cisplatin-induced apoptosis.