微小RNA-378转染对卵巢癌细胞株SKOV3增殖的影响及其机制

Effect of microRNA-378 transfection on proliferation of ovarian cancer cell line SKOV3

目的观察微小RNA-378(miRNA-378)转染对卵巢癌细胞株SKOV3增殖的影响,并探讨其可能机制。方法 miRNA-378过表达质粒转染的SKOV3细胞作为实验组,空白载体质粒转染的SKOV3细胞作为阴性对照组,SKOV3细胞作为空白对照组。采用Real-time PCR检测各组细胞中miRNA-378、环指蛋白31(RNF31)mRNA,Western blotting检测各组细胞中RNF31蛋白,CCK-8实验检测各组细胞增殖能力,克隆形成实验检测各组细胞克隆形成能力。结果实验组SKOV3细胞miRNA-378相对表达量高于阴性对照组和空白对照组(P均<0.05);实验组SKOV3细胞RNF31 mRNA和蛋白相对表达量分别低于阴性对照组和空白对照组(P均<0.05);实验组SKOV3细胞的增殖能力明显低于阴性对照组和空白对照组(P均<0.05);实验组SKOV3细胞的克隆形成能力低于阴性对照组和空白对照组(P均<0.05)。结论 miRNA-378转染可抑制卵巢癌细胞株SKOV3增殖,miRNA-378可能通过抑制RNF31基因表达而发挥抑制卵巢癌细胞增殖的作用。

Objective To observe the effect of microRNA-378 transfection on the proliferation of cancer cell line SKOV3 and to explore its possible mechanism.Methods We used the microRNA-378 overexpressing plasmid to transfect SKOV3 cells as the experimental group,and blank vector plasmid to transfect SKOV3 cells as the negative control group,meanwhile,the SKOV3 cells without transfection were taken as the blank control group.MicroRNA-378 and ring finger protein 31(RNF31)mRNA in each group was detected by real-time PCR,RNF31 protein in cells of each group was detected by Western blotting,the cell proliferation capacity in each group was detected by CCK-8,and colony formation assay was used to detect cell colony forming ability.Results The relative expression level of microRNA-378 in the SKOV3 cells of the experimental group was higher than that in the negative control group and the blank control group(P<0.05).The relative expression levels of RNF31 mRNA and protein in the experimental group was lower than that in the negative control group and the blank control group,respectively(P<0.05).The proliferation of SKOV3 cells in the experimental group was significantly lower than that in the negative control group and the blank control group(P<0.05).The cell clone formation ability in the experimental group was significantly lower than that the negative control group and the blank control group(P<0.05).Conclusion The microRNA-378 transfection can inhibit the cell proliferation of cancer cell line SKOV3,and microRNA-378 may inhibit the proliferation of ovarian cancer cells by inhibiting the expression of RNF31 gene.