miR-410对上皮性卵巢癌生物学行为影响及调控作用的实验研究

Experimental study on the effect and regulation of miR-410 on the biological behavior of epithelial ovarian cancer

目的:探讨miR-410对上皮性卵巢癌生物学功能的影响及调控作用。方法:检测上皮性卵巢癌组织和正常卵巢组织,以及卵巢癌SKOV3细胞和对照IOSE80细胞中miR-410表达水平。建立过表达miR-410瞬转SKOV3细胞体系,MTT法检测卵巢癌细胞增殖能力,Transwell法检测细胞侵袭能力,细胞划痕实验观察细胞迁移能力,Western blot法测定细胞中CCNB1蛋白表达水平。通过OncoLnc数据库评估卵巢癌患者的miR-410表达数据,分析miR-410对卵巢癌患者预后的影响。结果:卵巢癌组织中miR-410表达水平显著低于正常组织(P<0.05);SKOV3细胞中miR-410表达水平显著低于正常卵巢上皮IOSE80细胞(P<0.05)。转染后第2、3、4天,过表达miR-410组的SKOV3细胞增殖能力明显受到抑制(P<0.05);转染48h后,对照组和过表达组的穿膜细胞数分别为292.157±19.069、103.874±11.253,差异有统计学意义(P<0.05);划痕48h后,对照组与过表达miR-410组的迁移指数(MI)分别为(91.05±17.22)%和(59.36±11.29)%,差异有统计学意义(P<0.05)。转染miR-410后,SKOV3细胞中CCNB1蛋白表达水平显著下降(P<0.05)。miR-410低表达组卵巢癌患者与高表达组相比,总生存时间显著降低(P<0.05)。结论:上调miR-410表达可通过调控CCNB1抑制卵巢癌细胞的增殖与侵袭,低表达miR-410卵巢癌患者的总生存时间显著降低。

Objective:To investigate the effect of microRNA-410 on the biological function of epithelial ovarian cancer and its regulation.Methods:The expression of microRNA-410 in epithelial ovarian cancer tissues and normal ovarian tissues as well as ovarian cancer SKOV3 cells and control IOSE80 cells were detected by RT-PCR.To establish a transient SKOV3 cell system with over-expression of microRNA-410.MTT was used to detect the effect of microRNA-410 on the proliferation of epithelial ovarian cancer cells.To simulate the invasion of ovarian cancer cells through Transwell chamber.The ability of SKOV3 cells transfected with microRNA-410 to invade distantly was analyzed.The effect of overexpressed microRNA-410 on migration of SKOV3 cells was observed by cell scratch assay.After SKOV3 cells were transfected with microRNA-410,the expression of CCNB1 protein was measured.The expression of microRNA-410 in ovarian cancer patients was evaluated by OncoLnc database.To express data and analyze the effect of microRNA-410 on the prognosis of patients with ovarian cancer.Result:The expression level of microRNA-410 in ovarian cancer tissue was significantly lower than that in normal tissue(P<0.05).Compared with normal ovarian epithelial IOSE80 cells,the expression level of microRNA-410 in SKOV3 cells was significantly lower(P<0.05).The proliferation ability of SKOV3 cells in over-expressed microRNA-410 group was evident on the 2 nd,3 rd and 4 th day after transfection(P<0.05).After 48 hours of transfection,the number of perforating cells in the control group was higher than that in the experimental group(292.157±19.069 vs 103.874±11.253,P<0.05).After 48 hours of scratch,the MI of the control group and the overexpression group were(91.05±17.22)% and(59.36±11.29)%(P<0.05),respectively.CCNB1 protein expression level decreased(P<0.05).Compared with the high expression group,the total survival time of patients with ovarian cancer in the low expression group of microRNA-410 was significantly reduced(P<0.05).Conclusion:Up regulation of miR-410 expression can inhibit the proliferation,invasion and migration of ovarian cancer cells by regulating CCNB1.The overall survival time of ovarian cancer patients with low expression of miR-410 is significantly reduced.