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  

Effect of downregulated growth differential factor-9 on cell proliferative activity in mouse ovarian granulose cell

作　　者：马会明[1] 张永芳[1] 王蒙蒙[1] 李昀伽 蒲静[1] 于佳[1] 王燕蓉[1] 裴秀英[1] 徐仙[1] 

机构地区：[1]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,银川750004

出　　处：《安徽医科大学学报》2016年第1期51-55,共5页Acta Universitatis Medicinalis Anhui

基　　金：宁夏自然科学基金(编号:NZ11128)

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(P<0.05);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(P<0.01);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。Objective In the present study, we determined whether GDF9 siRNA were involved in the expressions of cyclin A, cyclin D1 and p27kipl which were key molecules in controlling cell cycling and cell proliferation by regulating cyclin D1 expression in mouse ovary granulose cell. Methods Real-time PCR and Western blot assay were used to investigate the expressions of cyclin D1, cyclin A, and p27kip1. We further observed the effect of GDF9 siRNA on the cell proliferation by regulating the expressions of cyclin D1, cyclin A, and p27kipl in mouse ovary granulose cell. Results We found that the expressions of cyclin D1 and cyclin A were down-regulated at both protein and mRNA levels in GDF9 siRNA group while p27kip1 was upregulated by real-time PCR and Western blot assay. Furthermore, GDF9 siRNA had additive effect in down-regulation of cyclin D1, cyclin A and upregulation of p27kip1. Compared with the control group, the expression level of cyelin D1 was significantly decreased（P 〈 0. 05） and p27kipl was significantly increased（P 〈 0. 05 ）. Conclusion These data suggest that cell cyclin signal path- way may coordinately regulate the cell proliferation and differentiation processes through up-regulating cyclin D1 and down-regulating p27kip1 of mouse ovary granulose cell.

关 键 词：GDF9 SIRNA 细胞增殖 细胞周期 卵巢颗粒细胞 小鼠 

分 类 号：R711.6[医药卫生—妇产科学]