微小RNA-185对前列腺癌细胞侵袭能力的影响及其机制

Effect of microRNA-185 on invasion ability of prostate cancer cells and its mechanism

目的观察前列腺癌(PC)细胞(DU145)中miR-185对活化素受体样激酶4(ALK4)基因的调控作用,及其对前列腺癌细胞侵袭能力的影响。方法收集河北省中医院泌尿外科2019年1月至2020年6月20例前列腺癌及其癌旁组织标本。采用荧光定量聚合酶链反应(PCR)方法检测前列腺癌组织中miR-185及ALK4基因的表达水平;将miR-185模拟物及阴性对照转染至DU145细胞中,使用双荧光素酶报告基因实验研究miR-185与ALK4基因间的关系;随后,将ALK4过表达质粒与miR-185模拟物共转染至DU145细胞中,采用Transwell细胞侵袭实验观察miR-185和ALK4表达水平对前列腺癌细胞侵袭的影响,两组间比较采用t检验。结果荧光定量PCR实验结果证明,miR-185在前列腺癌组织的表达水平(0.43±0.14)低于癌旁组织(1.03±0.06,t=6.736,P<0.05);相关性分析结果显示,miR-185与ALK4的表达呈负相关(r=-0.463,P<0.05);双荧光素酶报告基因分析实验表明,miR-185模拟物转染组荧光素酶活性(0.38±0.07)低于对照组(1.01±0.03,t=13.676,P<0.05);前列腺癌细胞中转染miR-185模拟物组侵袭细胞数[(125.33±13.01)个]显著低于对照组细胞数[(342.33±11.93)个,t=21.290,P<0.05];而与ALK4过表达质粒共转染后,miR-185模拟物转染组(322.00±18.33)与对照组(332.33±17.95)比较差异无统计学意义(t=0.698,P>0.05)。结论miR-185可通过靶向抑制ALK4基因抑制前列腺癌细胞侵袭能力。

Objective To observe the regulatory effect of microRNA(miR)-185 on activin receptor-like kinase 4(ALK4)gene in prostate cancer(PC)cells(DU145),and its influence on the invasion ability of PC cells.Methods Totally,20 specimens of PC and its adjacent tissues were collected from January 2019 to June 2020 in the Department of Urology,Traditional Chinese Medicine Hospital of Hebei Province.Real-time PCR was used to detect the expression levels of miR-185 and ALK4 genes in PC tissues.After transfecting miR-185 mimic and negative control into DU145 cells,dual luciferase reporter gene assay was used to observe the regulation of miR-185 on ALK4 gene.Subsequently,the overexpression plasmid of ALK4 and miR-185 mimics were cotransfected into DU145 cells.The effects of miR-185 and ALK4 expression levels on the invasion of PC cells were observed by Transwell assay.The t test was used for comparison between the two groups.Results Real-time PCR results proved that miR-185 expression(0.43±0.14)in PC tissues was significantly lower than that in adjacent tissues(1.03±0.06,t=6.736,P<0.05).Correlation analysis showed that miR-185 was negatively correlated with the expression of ALK4(r=-0.463,P<0.05).Dual luciferase reporter gene analysis showed that the luciferase activity in miR-185 mimic transfection group(0.38±0.07)was significantly lower than that in control group(1.01±0.03,t=13.676,P<0.05).The number of invasive cells in miR-185 mimic-transfected group[(125.33±13.01)cells]was significantly less than that in the control group[(342.33±11.93)cells,t=21.290,P<0.05].In addition,after co-transfection of miR-185 mimic with ALK4 overexpression plasmid,there was no significant difference in the number of invasive cells between the miR-185 mimic transfection group(322.00±18.33)and the negative control group(332.33±17.95).Conclusion MiR-185 can inhibit the invasion ability of PC cells by targeting the ALK4 gene.