微小RNA-23a调控人卵泡颗粒细胞凋亡的体外研究
杨晓庆;李莹;余兰;贾婵维;杨晓葵;
摘要(Abstract)：
目的:研究微小RNA-23a(miR-23a)对人卵泡颗粒细胞中蛋白激酶B(Akt)磷酸化和PTEN表达的影响,探讨miR-23a调控人颗粒细胞凋亡的分子机制。方法:分别将miR-23a前体(pre-miR-23a转染组)、miR-23a抑制剂(miR-23a抑制剂组)和pre-miRNA(阴性对照组)转染至体外培养的人卵泡颗粒细胞。应用实时定量荧光PCR技术检测miR-23a的表达,Western blot法检测Akt、p-Akt、PTEN、半胱天冬氨酸蛋白酶(Caspase-3)等凋亡相关蛋白的表达,并应用Hoechst 33258法检测细胞的凋亡率。结果:(1)与阴性对照组比较(4.05±0.92),转染pre-miR-23a后,miR-23a表达量(19.18±1.69)显著升高,转染miR-23a抑制剂后,miR-23a表达量(1.27±0.15)显著降低,差异均有统计学意义(P<0.05)。(2)转染pre-miR-23a后,Akt和p-Akt相对表达量显著低于阴性对照组(P<0.05);PTEN和Caspase-3裂解片断(Cleaved-caspase-3)相对表达量显著高于阴性对照组(P<0.05)。(3)pre-miR-23a转染组的细胞凋亡率为(17.87±2.20)%,显著高于阴性对照组(4.20±1.23)%(P<0.05)。结论:微小RNA-23a通过对其靶基因的调控参与PTEN/Akt信号通路介导的细胞凋亡,启动Caspase-3依赖的信号传导诱导体外培养人颗粒细胞的凋亡。
关键词(KeyWords)： 微小RNA-23a;蛋白激酶B;PTEN;颗粒细胞;凋亡
基金项目(Foundation): 国家自然科学基金项目(No:81170542);; 北京市卫生系统高层次人才培养项目(No:2011-3-071)
作者(Authors): 杨晓庆;李莹;余兰;贾婵维;杨晓葵;
DOI: 10.13283/j.cnki.xdfckjz.2014.03.017
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