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

miR-146a promotes the apoptosis of castration-resistant prostate cancer by inhibiting the expression of ROCK 1

目的:探讨mi RNA-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mi RNA-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mi RNA-146a对cleave-caspase 3表达的影响。构建ROCK1 3'UTR报告基因质粒,合成mi RNA-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mi R-146a是否与ROCK1相结合,采用免疫印迹法(Western blot)检测mi R-146a干扰后ROCK1蛋白量的表达。结果:Hoechst染色显示,转染mi RNA 146a后可以促进DU145和PC3的凋亡。高表达mi R-146a后cleave-caspase 3表达增加。Luciferase及Western blot显示,mi R-146a通过与靶基因ROCK1 3'UTR的结合抑制ROCK1的表达。结论:ROCK1是mi R-146a的靶基因,mi R-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 mi RNA-146 a promoting the apoptosis of castrationresistant prostate cancer cells（ DU145 / PC3）. Methods： Hoechst assay were performed to study the effect of high expression of mi RNA-146 a on apoptosis of prostate cancer cells. Western blot method were used to observe the high expression of mi RNA-146 a on expression of cleave-caspase 3. The ROCK1 3＇ UTR report gene plasmid was constructed and mi RNA-146 a minics was designed, followed by transferection into DU145 and PC3 cells respectively. The luciferase report gene detected the combination of mi RNA-146 a and its potential target genes ROCK1,Western blot was used to detect the amount of protein expression of ROCK1. Results： mi RNA-146 a promoted two kinds of prostate cancer cells DU145 and PC3 apoptosis. Cleave-caspase 3 expression was increased with high expression of mi RNA-146 a. The expression of ROCK1 was inhibited by combination of mi RNA-146 a with target gene ROCK1 3＇UTR. Conclusion： mi RNA-146 a promotes the apoptosis of prostate cancer cells by inhibiting the expression of ROCK1.