miR-152通过调控FOXR2表达抑制人前列腺癌细胞增殖的研究

miR-152 inhibiting the proliferation of human prostate cancer cells by regulating the expression of FOXR2

目的探讨人前列腺癌PC-3细胞中miR-152调控叉头框蛋白R2(FOXR2)对细胞增殖的影响。方法采用实时荧光定量PCR检测人正常前列腺上皮细胞RWPE-1和前列腺癌PC-3细胞中miR-152、FOXR2 mRNA的相对表达水平,Western blot检测FOXR2蛋白水平。miR-152 mimics、miR-152 inhibitor或FOXR2-siRNA转染PC-3细胞后,观察细胞增殖能力的变化。miR-152 mimics、miR-152 inhibitor分别与FOXR23′UTR双荧光素报告基因质粒共转染PC-3细胞,检测荧光素酶活性。miR-152 mimics和miR-152 inhibitor转染PC-3细胞后,检测miR-152、FOXR2 mRNA的相对表达水平及FOXR2蛋白水平。结果与RWPE-1细胞比较,PC-3细胞中miR-152的相对表达水平显著降低(P=0.001),而FOXR2 mRNA的相对表达水平及蛋白水平显著升高(P=0.003、0.003)。miR-152 mimics、miR-152 inhibitor或FOXR2-siRNA转染PC-3细胞后,细胞增殖能力分别显著降低、升高、降低(P=0.022、0.029、0.006)。miR-152 mimics和FOXR23′UTR野生型质粒共转染后荧光素酶活性被显著抑制(P=0.001),而共转染miR-152 inhibitor可显著增加荧光素酶活性(P=0.001)。与对照组比较,miR-152 mimics转染PC-3细胞后,miR-152的相对表达水平显著升高(P<0.001),而FOXR2 mRNA的相对表达水平及蛋白水平显著降低(P<0.001、0.001),细胞活性显著降低(P=0.013);miR-152 inhibitor转染PC-3细胞后,miR-152的相对表达水平显著降低(P<0.001),而FOXR2 mRNA的相对表达水平及蛋白水平显著升高(P<0.001、P=0.007),细胞活性显著升高(P=0.018)。结论miR-152可通过负调控FOXR2表达发挥抑制人前列腺癌PC-3细胞增殖的作用。

Objective To investigate the effect of microRNA(miR)-152 on cell proliferation via regulating fork-head box R2(FOXR2)in PC-3 cells of human prostate cancer cells.Methods The relative expression levels of miR-152 and FOXR2 mRNA in human normal prostate epithelial cell RWPE-1 cells and PC-3 cells were detected by qPCR,and the level of FOXR2 protein was detected by Western blot.After transfection of PC-3 cells with miR-152 mimics,miR-152 inhibitor or FOXR2-siRNA,the cell proliferation ability was observed.miR-152 mimics,miR-152 inhibitor and FOXR23′UTR double fluorescent reporter gene plasmid were co-transfected with PC-3 cells,and the luciferase activity was detected.After transfection of PC-3 cells with miR-152 mimics and miR-152 inhibitor,the relative expression levels of miR-152,FOXR2 mRNA and the level of FOXR2 protein were detected.Results Compared with RWPE-1 cells,the relative expression level of miR-152 in PC-3 cells was significantly decreased(P=0.001),while the relative expression level of FOXR2 mRNA and protein level was significantly increased(P=0.003,0.003).After transfection of PC-3 cells with miR-152 mimics,miR-152 inhibitor or FOXR2-siRNA,cell proliferation ability was significantly decreased,increased and decreased respectively(P=0.022,0.029,0.006).Co-transfection of miR-152 mimics and FOXR23′UTR wild type plasmids significantly inhibited the luciferase activity(P=0.001),while Co-transfection of miR-152 inhibitor significantly increased the luciferase activity(P=0.001).Compared with the control group,the relative expression level of miR-152 was significantly increased after miR-152 mimics transfected into PC-3 cells(P<0.001),while the relative expression level of FOXR2 mRNA and protein level was significantly decreased(P<0.001,0.001)and the cell viability was significantly decreased(P=0.013).The relative expression level of miR-152 was significantly decreased(P<0.001)after miR-152 inhibitor transfected into PC-3 cells,while the relative expression level of FOXR2 mRNA and protein was significantly increased(P<0.001,P=0.007)and the cell viability was significantly increased(P=0.018).Conclusion miR-152 can inhibit the proliferation of human prostate cancer PC-3 cells by negatively regulating FOXR2 expression.