赵贵旭王倩倩孙姗姗顾红琴席晓薇△ 张箴波丰有吉.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[J].,2012,12(8):1405-1409 |
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 |
Effects of Follicle Stimulating Hormone on Proliferation, Apoptopsis andInvasion of Immortalized Human Ovarian EpithelialCells Through OCT4 Gene |
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DOI: |
中文关键词: OCT4 卵泡刺激素 卵巢上皮细胞 增殖 凋亡 侵袭 |
英文关键词: OCT4 Follicle stimulating hormone Ovarian epithelial cells Proliferation Apoptpsis Invasion |
基金项目:国家自然科学基金项目(81172478) |
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中文摘要: |
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;③
FSH+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。 |
英文摘要: |
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. |
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