miR-21调节P53的表达及对子宫内膜癌Ishikawa细胞增殖、凋亡的影响  

Effects of miR-21 on expression of P53 and its effects on proliferation and apoptosis of endometrial carcinoma cells in vitro

作　　者：王淑芳 底建敏[2] 赵向寨[2] 白云[2] 冯玉珍[2] WANG Shufang;DI Jianmin;ZHAO Xiangzhai(Hospital for Maternal and Child Health of Qinhuangdao City,Hebei,Qinhuangdao 066000,China)

机构地区：[1]河北省秦皇岛市妇幼保健院,066000 [2]河北医科大学第三医院妇产科

出　　处：《河北医药》2018年第6期866-869,873,共5页Hebei Medical Journal

基　　金：河北省医学科研究重点课题(编号:20170661)

摘　　要：目的探讨子宫内膜癌Ishikawa细胞中miR-21的表达及其对Ishikawa细胞中p53蛋白表达和细胞增殖、凋亡的影响。方法脂质体介导将miR-21的特异性抑制剂反义寡核苷酸转染到Ishikawa细胞为实验组,转染不相关序列、脂质体和未转染组细胞为阴性对照组、脂质体对照组和空白对照组。CCK-8法和流式细胞学法检测细胞增殖和凋亡情况;实时定量PCR检测miR-21和P53mRNA的表达水平;Western blot检测P53蛋白表达。结果激光共聚焦显微镜观察,实验组及阴性对照组荧光表达均强于其他组,证明在本次试验中质粒转染成功。CCK-8法检测发现4组细胞整体呈增殖状态,空白对照组、脂质体对照组和实验组增殖速度稍快,阴性对照组最慢,各组之间比较差异无统计学意义(P>0.05)。细胞转染3 d后,实验组与其他3组比较早期凋亡所占的比例明显偏低(P<0.05),晚期凋亡细胞所占比例相似(P>0.05)。实验组miR-21的表达量相对于各对照组明显降低(P<0.05),实验组和阴性对照组P53mRNA的表达量相对于空白对照组和脂质体对照组均有明显降低(P<0.05)。实验组p53蛋白表达明显高于其他对照组,与各组间比较差异有统计学意义(P<0.05)。结论 miR-21能够促进子宫内膜癌Ishikawa细胞凋亡,对P53基因的调控作用不明显,但能在转录后水平抑制P53蛋白的表达,可能作为子宫内膜癌基因治疗的新靶点。Objective To investigate the effects of miR-21 on expression of P53 and its effects on proliferation and apoptosis of endometrial carcinoma Ishikawa cells.Methods Through the mediation of lipofectamine TM2000 liposome transfection reagent,the endometrial carcinoma-Ishikawa cells were transfected with miR-21 specific inhibitor antisense oligonucleotides(experimental group),and the cells that were transfected with irrelevant miR-21 specific inhibitor nucleotide sequence or liposome were served as negative control group and liposome control group,respectively.Moreover the cells without transfection were served as blank control group.The cell growth activity and cell apoptosis were detected by CCK-8,flow cytometry,respectively.The expression levels of miR-21 and P53 were detected by Real-time quantitative RT-PCR,and the expression levels of P53 protein were measured by Western Blot.Results The laser confocal microscopy showed that the fluorescent expression levels in experimental group and negative control group were higher than those in the other groups,which confirmed that plasmid transfection was successful in this study.The CCK-8 experiments found that the cells presented proliferation state in the four groups,however,the proliferation speed in blank control group,liposome control group and experimental group was slightly faster,which in negative control group was the slowest,however,there was no significant difference between two groups each other(P>0.05).At 3d after cell transfection,the proportion of early apoptosis cells in experimental group was significantly lower than that in the other three groups(P<0.05),but there was no significant difference in the proportion of non-viable apoptotic cell among groups(P>0.05).Moreover the expression levels of miR-21 in experimental group were significantly increased,as compared with those in the other three groups(P<0.05).However the expression levels of miR-21 in experimental group were significantly lower than those in the other groups(P<0.05).As compared with those i

关 键 词：子宫内膜癌 MICRORNA MIR-21 P53 ISHIKAWA 

分 类 号：R737.33[医药卫生—肿瘤]