miR-21对化疗药物诱导卵巢颗粒细胞凋亡的影响及其机制  

miR-21 rescues the apoptosis of chemotherapy-induced ovarian granulosa cells

作　　者：汪庆如 陈小莹[1] 李欣然[1] 王清[1] 付霞霏[1] WANG Qing - ru, CHEN Xiao - ying, LI Xin - ran, WANG Qing, FU Xia -fei.(Department of Obstetrics and Gynecology, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, Guangdong, Chin)

机构地区：[1]南方医科大学珠江医院妇产科,广东广州510282

出　　处：《广东医学》2018年第18期2709-2712,共4页Guangdong Medical Journal

基　　金：国家自然科学基金青年科学基金资助项目(编号:81300462)

摘　　要：目的探讨慢病毒介导miR-21对化疗诱导卵巢颗粒细胞凋亡的影响及其机制。方法采用分子生物学方法构建miR-21慢病毒表达载体(LV-miR-21),分离培养大鼠卵巢颗粒细胞。分为正常组、磷酰胺氮芥(PM)组、miR-21组。正常组卵巢颗粒细胞培养基中不加PM,PM组加入30μmol/L的PM诱导卵巢颗粒细胞凋亡,miR-21组卵巢颗粒细胞培养基中加PM诱导凋亡24 h,随后转染LV-miR-21。48 h后采用流式细胞仪检测法检测3组卵巢颗粒细胞凋亡率,用荧光定量PCR及Western blot法检测同源丢失性磷酸酶张力蛋白基因(PTEN)、程序性死亡蛋白4(PDCD4)的mRNA及蛋白质表达水平。结果正常组、PM组、miR-21组的凋亡率差异有统计学意义(F=78. 00,P=0. 00),miR-21组凋亡率低于PM组(P <0. 05),但仍高于正常组(P <0. 05)。3组的miR-21表达水平差异有统计学意义(F=1 226. 35,P=0. 00),miR-21组的miR-21表达水平显著高于正常组及PM组(P <0. 05)。3组PTEN的mRNA和蛋白质表达水平差异均有统计学意义(F=90. 37,P=0. 00; F=44. 34,P=0. 00),miR-21组的表达水平显著低于PM组(P <0. 05)。3组PDCD4的mRNA和蛋白表达水平差异均有统计学意义(F=198. 59,P=0. 00; F=60. 25,P=0. 00),miR-21组的表达水平显著低于PM组(P <0. 05)。结论 miR-21通过下调靶基因PTEN、PDCD4的表达,抑制化疗药物诱导的卵巢颗粒细胞凋亡。Objective To study the effects of miR-21 on chemotherapy-induced apoptosis of ovarian granulosa cells（ GCs）. Methods GCs were isolated in vitro and divided into normal group（ without PM and transfection）,PM group（ with PM but no transfection） and miR-21 group（ with PM and transfected with miR-21 lentiviral vector）. GCs were transfected with miR-21 lentiviral vector（ LV-miR-21） to obtain miR-21-expressing GCs. Apoptosis was induced by phosphamide mustard（ PM）. The apoptosis rate,the miR-21,and the mRNA and protein expression of target genes,including phosphatase and tensin homolog deleted on chromosome ten（ PTEN） and programmed cell death 4（ PDCD4）,were assessed in GCs. Results The apoptosis rates of normal group,PM group and miR-21 group were statistically different（ F = 78. 00,P = 0. 00）; which was significantly lower in miR-21 group than PM group,but still higher than normal group. The expression levels of miR-21 in these three groups were statistically different（ F = 1 226. 35,P =0. 00）; which was significantly higher in miR-21 than those in normal group and PM group（ P〈 0. 05）. The expression levels of PDCD4 mRNA and protein in these three groups were statistically different（ F = 90. 37,P = 0. 00; F = 44. 34,P = 0. 00）; which were significantly lower in miR-21 group than PM group（ P〈 0. 05）. The expression levels of PTEN mRNA and protein in these three groups were statistically different（ F = 198. 59,P = 0. 00; F = 60. 25,P = 0. 00）;which was significantly lower in miR-21 than PM group（ P〈 0. 05）. Conclusions The expression of miR-21 in GCs rescues the apoptosis of chemotherapy-induced apoptosis of ovarian granulosa cells via PTEN and PDCD4.

关 键 词：MIRNA-21 卵巢颗粒细胞 化疗 PTEN PDCD4 

分 类 号：R711.75[医药卫生—妇产科学]