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miR-146a通过抑制ROCK1基因的表达促进去势抵抗性前列腺癌细胞的凋亡
作者:蒋亮1  刘春辉1  许斌2  陈明2 
单位:1. 东南大学 医学院, 江苏 南京 210009;
2. 东南大学 附属中大医院, 江苏 南京 210009
关键词:前列腺癌 Rho相关卷曲螺旋蛋白激酶1 细胞凋亡 
分类号:R737.25
出版年·卷·期(页码):2015·34·第三期(357-360)
摘要:

目的:探讨miRNA-146a对激素非依赖前列腺癌细胞DU145、PC3凋亡的影响极其作用机制.方法:通过Hoechst染色方法观察高表达miRNA-146a对前列腺癌细胞凋亡的影响.通过免疫印迹法(Western blot)观察高表达miRNA-146a对cleave-caspase 3表达的影响.构建ROCK1 3'UTR报告基因质粒,合成miRNA-146a minics,分别共转染DU145、PC3两种前列腺癌细胞,采用荧光素酶报告基因(Luciferase)实验检测miR-146a是否与ROCK1相结合,采用免疫印迹法(Western blot)检测miR-146a干扰后ROCK1蛋白量的表达.结果: Hoechst染色显示,转染miRNA 146a后可以促进DU145和PC3的凋亡.高表达miR-146a后cleave-caspase 3 表达增加.Luciferase及Western blot显示,miR-146a通过与靶基因ROCK1 3'UTR的结合抑制ROCK1的表达.结论:ROCK1是miR-146a的靶基因,miR-146a通过抑制ROCK1基因的表达促进前列腺细胞的凋亡.

Objective: To explore the affect and mechnism of miRNA-146a promoting the apoptosis of castration-resistant prostate cancer cells (DU145/PC3). Methods: Hoechst assay were performed to study the effect of high expression of miRNA-146a on apoptosis of prostate cancer cells. Western blot method were used to observe the high expression of miRNA-146a on expression of cleave-caspase 3. The ROCK1 3'UTR report gene plasmid was constructed and miRNA-146a minics was designed, followed by transferection into DU145 and PC3 cells respectively. The luciferase report gene detected the combination of miRNA-146a and its potential target genes ROCK1, Western blot was used to detect the amount of protein expression of ROCK1. Results: miRNA-146a promoted two kinds of prostate cancer cells DU145 and PC3 apoptosis. Cleave-caspase 3 expression was increased with high expression of miRNA-146a. The expression of ROCK1 was inhibited by combination of miRNA-146a with target gene ROCK1 3'UTR. Conclusion: miRNA-146a promotes the apoptosis of prostate cancer cells by inhibiting the expression of ROCK1.

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