miR-613通过下调Wnt/β-catenin信号通路活性抑制前列腺癌细胞的增殖与侵袭

miR-613 inhibits the proliferation and invasion of prostate cancer cells by down-regulating the activity of Wnt/β-catenin signaling pathway

目的:研究miR-613在人前列腺癌组织中的表达情况,探讨miR-613是否通过下调Wnt信号通路活性抑制前列腺癌细胞系细胞的增殖和侵袭能力。方法:收集临床前列腺癌组织及配对癌旁组织20例,通过实时荧光定量PCR(RT-qPCR)检测各组组织中miR-613的表达情况。进一步在细胞实验中,通过转染miR-613 mimic和miR-NC至离体培养的PC-3、DU-145细胞中,随后,采用MTT法、平板克隆实验检测细胞增殖和Matrigel侵袭实验测定前列腺癌细胞的侵袭情况,采用荧光素酶分析方法评估Wnt信号通路活性变化,采用实时荧光定量PCR(RT-qPCR)检测Wnt/β-catenin信号通路下游靶基因的转录(包括Cyclin D1和c-Myc),WB法检测细胞中β-catenin、c-Myc和Cyclin D1的表达量。结果:相比配对癌旁组织,miR-613在前列腺癌组织中的表达降低(P<0.01);在体外细胞实验中,相比于miR-NC组,转染miR-613 mimic后,PC-3、DU-145细胞增殖能力下降(P<0.05),PC-3、DU-145细胞的迁移侵袭能力下降(P<0.01);miR-613的过表达显著降低Wnt信号通路活性、β-catenin蛋白表达及Wnt信号下游靶基因Cyclin D1和c-Myc的转录及蛋白表达。结论:miR-613通过抑制Wnt/β-catenin信号通路来影响前列腺癌细胞的增殖与侵袭,为前列腺癌的潜在治疗靶点之一。

Objective:To investigate the expression of miR-613 in human prostate cancer tissues and to investigate whether miR-613 down-regulate the proliferation and invasion ability of prostate cancer cell lines by inhibiting the activity of Wnt signaling pathway.Methods:Twenty clinical prostate cancer tissues and matched adjacent tissues were collected,and the expression of miR-613 in each group was detected by real-time fluorescence quantitative PCR(RT-qPCR).Further in cell experiments,miR-613 mimic and miR-NC were transfected into PC-3 and DU-145 cells cultured in vitro.MTT assay,plate cloning assay detected cell proliferation,and Matrigel invasion assay detected prostate cancer cell invasion.Changes in Wnt signaling pathway activity were evaluated by luciferase assays.Real-time fluorescence quantitative PCR(RT-qPCR)detected transcription of target genes downstream of Wnt/β-catenin signaling pathway(including Cyclin D1 and c-Myc).The expressions ofβ-catenin,c-Myc and Cyclin D1 were detected by WB method.Results:Compared with the matched paracancerous tissues,the expression of miR-613 was decreased in prostate cancer tissues(P<0.01).In vitro cell experiments,compared with miR-NC group,after transfecting miR-613 mimic,the cell proliferation capacity of PC-3 and DU-145 cells decreased(P<0.05),and the migration and invasion ability of PC-3 and DU-145 cells decreased(P<0.01).Overexpression of miR-613 significantly reduced Wnt signaling pathway activity,β-catenin protein expression,and transcription and protein expression of Cyclin D1 and c-Myc,the downstream target genes of Wnt signaling.Conclusion:miR-613 affects the proliferation and invasion of prostate cancer cells by inhibiting Wnt/β-catenin signaling pathway,and is one of the potential therapeutic targets for prostate cancer.