miR-29a过表达对人前列腺癌细胞凋亡的调控机制研究  

STUDY ON THE REGULATORY MECHANISM OF MIR-29A OVEREXPRESSION ON APOPTOSIS OF HUMAN PROSTATE CANCER CELLS

作　　者：罗于杰 蓝建华[1] 黄国华[1] LUO Yujie;LAN Jianhua;HUANG Guohua(Department of Urology,Guang’an People’s Hospital,Sichuan 638000,China)

机构地区：[1]四川省广安市人民医院泌尿外科,638000

出　　处：《中国煤炭工业医学杂志》2021年第1期15-20,共6页Chinese Journal of Coal Industry Medicine

基　　金：四川省科技厅科技支撑计划项目(编号:2018ZC0421)。

摘　　要：目的探讨过表达微型RNA-29a(miR-29a)对人前列腺癌(PCa)细胞凋亡的影响及其作用机制。方法体外培养人前列腺癌细胞,并分为空白对照组、miR-29a拟似物转染阴性对照组(NC组)、miR-29a拟似物转染组(miR-29a组),阴性对照组使用空白质粒转染,miR-29a拟似物转染组使用含有目的基因的miR-29a进行转染,转染48 h后用荧光定量PCR方法检测人前列腺癌细胞miR-29a的表达变化,采用MTT法检测人前列腺癌细胞生长情况,使用流式细胞仪检测细胞凋亡率及周期变化,Western blot法检测B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-2X相关的蛋白质(Bax)、含半胱氨酸的天冬氨酸蛋白水解酶-3(Caspase-3)、含半胱氨酸的天冬氨酸蛋白水解酶-9(Caspase-9)、磷酸化蛋白激酶(P-Akt)和蛋白激酶(Akt)蛋白表达量。结果miR-29a组miR-29a的表达高于空白对照组和NC组(均P<0.05),NC组与空白对照组miR-29a表达水平差异无统计学意义(P>0.05)。MTT结果显示miR-29a组细胞生长抑制率均高于空白对照组和NC组(均P<0.05),NC组与空白对照组差异无统计学意义(P>0.05)。流式细胞术检验结果显示空白对照组、NC组和miR-29a组细胞凋亡率依次升高(P<0.05)。与空白对照组和NC组比较,miR-29a组人PCa细胞中G1期细胞比例增加,S期细胞比例减少(均P<0.05),G2期细胞比例无明显变化(P>0.05)。Western blot结果显示与空白对照组和NC组比较,Caspase-3、Caspase-9、Bax蛋白表达水平升高,Bcl-2、P-Akt蛋白表达水平降低(均P<0.05),Akt蛋白表达水平无明显变化(P>0.05)。结论过表达miR-29a可以通过下调抑凋亡蛋白Bcl-2,上调促凋亡蛋白Bax、凋亡调控因子Caspase-3和Caspase-9蛋白表达,抑制PI3K/AKT信号通路达到抑制前列腺癌细胞增殖,促进前列腺癌细胞凋亡的效果。Objective To explore the effects and role mechanism of microRNA-29 a(miR-29 a)overexpression on apoptosis of human prostate cancer(PCa)cells.Methods Human prostate cancer cells were cultured invitro and divided into blank control group,miR-29 a mimic negative control group(NC group)and miR-29 a mimic transfection group(miR-29 a group).NC group was transfected with a blank plasmid,and miR-29 a group was transfected with miR-29 a containing the target gene.After 48 h of transfection,the expression of human prostate cancer cell miR-29 a was detected by fluorescent quantitative PCR.The human prostate cancer cell growth was detected by MTT assay.The apoptosis rate and periodic changes were detected by flow cytometry.The protein expression levels of B-cell lymphoma 2(Bcl-2),B lymphoma-2 X-related protein(Bax),Cysteinyl aspartate specific proteinase-3(Caspase-3),Cysteinyl aspartate specific proteinase-9(Caspase-9),phosphorylated protein kinase(P-Akt)and protein kinase(Akt)were detected by Western blot.Results The expression of miR-29 a in miR-29 a group was significantly higher than that in blank control group and NC group(P<0.05),and the difference between NC group and blank control group was not significant in miR-29 a expression(P>0.05).MTT results showed that the cell growth inhibition rate in miR-29 a group was significantly higher than that in blank control group and NC group(P<0.05),and the difference between NC group and blank control group was not significant(P>0.05).The results of flow cytometry showed that the apoptosis rate in blank control group,NC group and miR-29 a group was increased in turn(P<0.05).Compared with blank control group and NC group,the proportion of cells in G1 phase of human PCa cells in miR-29 a group was significantly increased while the proportion of cells in S phase was significantly decreased(P<0.05),and the proportion of cells in G2 phase was not significantly changed(P>0.05).Western blot analysis showed that compared with blank control group and NC group,the protein expression level

关 键 词：前列腺癌细胞 miR-29a 细胞凋亡 

分 类 号：R737.25[医药卫生—肿瘤]