OCT4 介导卵泡刺激素对永生化人卵巢上皮细胞株增殖、 凋亡和侵袭能力的影响

目的:探讨OCT4基因对卵泡刺激素作用下的永生化人卵巢上皮细胞株(Moody细胞)增殖、凋亡和侵袭能力影响。方法:将不同浓度的FSH(0、25、50、100mIU/ml)作用于Moody细胞48小时,应用Western-blot技术检测OCT4表达情况。采用慢病毒介导将重组质粒OCT4稳定转染至人卵巢上皮细胞株中,应用Western-blot法鉴定OCT4蛋白表达情况。FSH以50 mIU/ml作为工作浓度,实验对象分为4组:①siCon组,转染空载体的阴性对照组;②OCT4组:稳定转染OCT4基因的Moody细胞组;③FSH+siCon组:以FSH处理的siCon组;④FSH+OCT4组:以FSH处理的OCT4组。采用MTT比色法检测各组细胞的增殖情况,流式细胞仪检测各组细胞凋亡情况,Transwell侵袭实验检测各组细胞侵袭能力的变化。结果:(1)随着FSH浓度的增加,Moody细胞中OCT4蛋白表达逐渐增高,在FSH浓度为50 mIU/ml时达最高;(2)OCT4基因成功转染至Moody细胞中,经Western-blot检测该基因在细胞中进行蛋白高表达;(3)FSH+OCT4组细胞增殖活性明显增高,同时凋亡率降低,与另外三组相比差异具有统计学意义(P<0.05);(4)在FSH作用下,转染OCT4后明显增强了细胞的侵袭能力,与另外三组相比差异具有统计学意义(P<0.05)。结论:OCT4介导了FSH对人卵巢上皮细胞增殖、凋亡、侵袭活性的调控。

Effects of Follicle Stimulating Hormone on Proliferation, Apoptopsis and Invasion of Immortalized Human Ovarian Epithelial Cells Through OCT4 Gene

Objective: To investigate the effects of follicle stimulating hormone(FSH) on the proliferation,apoptosis,and invasion of immortalized ovarian epithelial cells(Moody) through OCT4 gene.Methods: The Moody cells were stimulated by FSH with the concentrations of 0,25,50,100 mIU/ml for 48h.The expression of OCT4 protein was detected by Western bloting.The recombinant plasmids were stably transfected into human ovarian epithelial cells by Lentiviral vector.Western bloting was used to detect the expression of OCT4 protein.And then the cells were stimulated with 50 mIU/ml FSH and were divided into 4 groups: ①siCon: si-Negative control transfected with empty vecot;②OCT4: Moody cells transfected with OCT4;③FSH+siCon;siCon group treated by FSH;④FSH+ OCT4: OCT4 group treated by FSH.The proliferation effects of the cells were detected by MTT assay,the apoptosis were examined by flow cytometry and the invasive and migrated ability were examined by Transwell assay.Results: The protein expression level of OCT4 treated with FSH of the concentration of 25~100 mIU/ml were all significantly higher than those without FSH treatment and was highest with the concentration of 50 mIU/ml.OCT4 gene was transfected into the Moody cells and Western blotting analysis revealed that the expression of OCT4 protein in stable transfeted cells.The proliferation effects were significantly higher in the FSH+ OCT4 group and apoptosis rates were significantly lower in the FSH+ OCT4 group compared with other groups respectively(P<0.05).The number of invaded Moody cells was significantly higher in the FSH+ OCT4 group than other groups.Conclusion: OCT4 plays an important role in mediating FSH-dependent proliferation,apoptpsis,invasion of human ovarian epithelial cells.