生长分化因子-9下调对小鼠卵巢颗粒细胞增殖能力的影响

发布时间：2018-02-06 03:30

本文关键词： GDF siRNA 细胞增殖细胞周期卵巢颗粒细胞小鼠　出处：《安徽医科大学学报》2016年01期 　论文类型：期刊论文

【摘要】：目的探讨生长分化因子-9(GDF9)对小鼠卵巢颗粒细胞增殖能力基因周期蛋白A(cyclin A)、周期蛋白D1(cyclin D1)和周期蛋白依赖性激酶抑制因子27(p27kip1)表达的影响。方法利用CCK-8检测法检测细胞的增殖能力;使用流式细胞术、Real-time PCR和Western blot法验证GDF9 siRNA的干扰效率;采用Real-time PCR和Western blot法检测细胞增殖cyclin A、cyclin D1和p27kip1 mRNA和蛋白表达情况。结果 GDF9 siRNA显著抑制颗粒细胞增殖活力,其中72 h抑制最明显(P0.05);Real-time PCR和Western blot法检测结果显示GDF9 siRNA显著下调小鼠卵巢颗粒细胞中GDF9 mRNA及蛋白的表达(P0.01);Realtime PCR法检测结果显示,细胞cyclin A、cyclin D1 mRNA表达降低,p27kip1 mRNA表达增强;Western blot法结果显示,GDF9 siRNA可促进细胞周期蛋白依赖性激酶抑制剂p27kip1的表达,同时下调G2/M期调控蛋白cyclin A、cyclin D1的表达。结论靶向抑制小鼠卵巢颗粒细胞中GDF9的表达,下调细胞cyclin A、cyclin D1的表达,上调P27 kip1的表达,显著抑制细胞的增殖。
[Abstract]:Objective to investigate the effect of growth differentiation factor-9 (GDF9) on ovarian granulosa cell proliferation ability gene cyclin A (cyclin A). The expression of cyclin D1 (cyclin D1) and cyclin dependent kinase inhibitor (27p27kip1) were detected by CCK-8 assay. Real-time PCR and Western blot were used to verify the interference efficiency of GDF9 siRNA. The proliferation of cyclin A was detected by Real-time PCR and Western blot. Expression of cyclin D1 and p27kip1 mRNA and protein. Results GDF9 siRNA significantly inhibited the proliferation of granulosa cells. In 72 h, the inhibition of P0.05 was the most obvious. Real-time PCR and Western blot assay showed that GDF9 siRNA significantly down-regulated GDF9 in mouse ovarian granulosa cells. Expression of mRNA and protein (. P0.01; Realtime PCR assay showed that the expression of cyclin cyclin D1 mRNA decreased and p27kip1 mRNA expression increased. The results of Western blot assay showed that GDF9 siRNA could promote the expression of p27kip1, a cyclin dependent kinase inhibitor. At the same time, it down-regulated the expression of G _ 2 / M phase regulatory protein cyclin Ancyclin D1. Conclusion the expression of GDF9 in mouse ovarian granulosa cells is inhibited by targeting. The expression of cyclin cyclin D1 was down-regulated, and the expression of P27 kip1 was up-regulated, which significantly inhibited cell proliferation.
【作者单位】：宁夏医科大学生育力保持教育部重点实验室宁夏回族自治区生殖与遗传重点实验室人体解剖与组织胚胎学系宁夏医科大学总医院生殖医学中心;
【基金】：宁夏自然科学基金(编号:NZ11128)
【分类号】：R321.1

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