circFoxo3表达对卵巢癌细胞增殖、凋亡和转移的作用机制研究

Study on the mechanism of circFoxo3 expression on the proliferation,apoptosis and metastasis of ovarian cancer cells

目的研究环状RNA叉头盒O3(circFoxo3)对卵巢癌细胞增殖、凋亡和转移能力的影响,并探讨其发挥作用的可能机制。方法常规培养卵巢癌细胞系CoC1、SKOV3、CAOV4和正常卵巢上皮细胞IOSE80,qRT-PCR检测circFoxo3在卵巢癌细胞系和正常卵巢上皮细胞中的表达水平。选择高表达circFoxo3的卵巢癌细胞转染circFoxo3敲减质粒,分为si-NC和si-circFoxo3组。MTS和平板克隆实验检测circFoxo3对卵巢癌细胞增殖能力的影响;流式细胞仪检测circFoxo3对卵巢癌细胞凋亡的影响;Transwell实验检测circFoxo3对卵巢癌细胞转移能力的影响;Western blotting检测circFoxo3对PI3K/AKT信号通路相关蛋白PI3K、pPI3K、AKT和pAKT表达的影响。结果circFoxo3在卵巢癌细胞中的表达显著高于在正常卵巢上皮细胞IOSE80中的表达(P<0.05);与si-NC组相比,si-circFoxo3组细胞的增殖能力降低,细胞凋亡增加,细胞转移能力降低;si-circFoxo3组细胞pPI3K和pAKT蛋白表达降低。结论circFoxo3在卵巢癌细胞中表达上调,可能通过调控PI3K/AKT促进卵巢癌细胞的进展。circFoxo3可能是治疗卵巢癌的潜在靶点。

Objective To study the effects of circular RNA Forkhead box O3(circFoxo3)on the proliferation,apoptosis and metastasis of ovarian cancer cells,and to explore the possible mechanism of its function.Methods Routinely culture the expression of ovarian cancer cell lines CoC1,SKOV3,CAOV4 and normal ovarian epithelial cells IOSE80.qRT-PCR was used to detect the expression level of circFoxo3 in ovarian cancer cell lines.Ovarian cancer cells with high expression of circFoxo3 were transfected with circFoxo3 knockdown plasmid,and they were divided into si-NC and si-circFoxo3 groups.MTS and plate cloning experiments were used to detect the effect of circFoxo3 on the proliferation of ovarian cancer cells;flow cytometry was used to detect the effect of circFoxo3 on the apoptosis of ovarian cancer cells;Transwell experiment was used to detect the effect of circFoxo3 on the metastasis of ovarian cancer cells;western blotting was used to detect the effect of circFoxo3 on PI3K/AKT signaling pathway related proteins PI3K,pPI3K,AKT and pAKT expression.Results The expression of circFoxo3 in ovarian cancer cells was significantly higher than that in normal ovarian epithelial cells IOSE80(P<0.05);compared with the si-NC group,the proliferation ability of the si-circFoxo3 group was reduced,and the cell apoptosis was increased,cell transfer ability was decreased;pPI3K and pAKT protein expression were decreased in si-circFoxo3 group cells.Conclusion circFoxo3 is up-regulated in ovarian cancer cells,and may promote the progression of ovarian cancer cells by regulating PI3K/AKT.circFoxo3 may be a potential target for the treatment of ovarian cancer.