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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。
Objective: To investigate the effect of OCT4 gene on the proliferation, apoptosis and invasion of immortalized human ovarian epithelial cell line (Moody’s cell line) induced by follicle-stimulating hormone. Methods: Moody cells were treated with different concentrations of FSH (0, 25, 50, 100 mIU / ml) for 48 hours. The expression of OCT4 was detected by Western-blot. The lentivirus-mediated recombinant plasmid OCT4 stably transfected into human ovarian epithelial cell line, the use of Western-blot to identify OCT4 protein expression. FSH at 50 mIU / ml as the working concentration, the experimental subjects were divided into four groups: ①siCon group, empty vector transfected negative control group; ② OCT4 group: stably transfected OCT4 gene Moody cell group; ③FSH + siCon group: FSH treatment Of siCon group; ④ FSH + OCT4 group: FSH-treated OCT4 group. MTT assay was used to detect the proliferation of each group. Flow cytometry was used to detect the apoptosis of each group. Transwell invasion assay was used to detect the changes of cell invasiveness in each group. Results: (1) With the increase of FSH concentration, the expression of OCT4 protein in Moody cells gradually increased, reaching the highest at FSH concentration of 50 mIU / ml; (2) OCT4 gene was successfully transfected into Moody cells, (3) The proliferation activity of FSH + OCT4 group was significantly higher than that of the other three groups (P <0.05); (4) Under the action of FSH, the transfection of OCT4 significantly enhanced cell invasion, compared with the other three groups, the difference was statistically significant (P <0.05). Conclusion: OCT4 mediates the regulation of FSH on the proliferation, apoptosis and invasion of human ovarian epithelial cells.