干扰LINC00308促进miR-634对前列腺癌细胞增殖与凋亡的影响  

Effect of interfering with LINC00308 to promote miR-634 on proliferation and apoptosis of prostate cancer cells

作　　者：魏仁波[1] 张唯力[2] 卓晖[1] 杨镒魟[3] 熊黎强[1] 杨鹏[4] 魏武然[5] 曹敏[1] WEI Renbo;ZHANG Weili;ZHUO Hui;YANG Yihong;XIONG Liqiang;YANG Peng;WEI Wuran;CAO Min(Department of Urology, Chengdu Third People’s Hospital, Chengdu 610031, China;Department of Urology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China;Department of Andrology, West China Second Hospital, Sichuan University, Chengdu 610041, China;Department of Pathology, Chengdu Third People’s Hospital, Chengdu 610031, China;Department of Urology, West China Hospital, Sichuan University, Chengdu 610041, China)

机构地区：[1]成都市第三人民医院泌尿外科,四川成都610031 [2]重庆医科大学附属第二医院泌尿外科,重庆400010 [3]四川大学华西第二医院男科,四川成都610041 [4]成都市第三人民医院病理科,四川成都610031 [5]四川大学华西医院泌尿外科,四川成都610041

出　　处：《西部医学》2021年第2期191-197,204,共8页Medical Journal of West China

基　　金：四川省卫生健康委员会科研课题(18PJ461)。

摘　　要：目的研究干扰长链非编码RNA(lncRNA)LINC00308是否通过促进微小RNA(miRNA)-634影响前列腺癌细胞的增殖与凋亡。方法实时荧光定量PCR(qRT-PCR)检测前列腺癌组织中LINC00308的表达。利用LINC00308小干扰RNA(si-LINC00308)转染前列腺癌细胞PC-3M和DU145,构建干扰LINC00308的细胞,采用qRT-PCR检测LINC00308和miR-634的表达水平,流式细胞术评估细胞的周期与凋亡,蛋白质印迹法(Western Blot)检测细胞核相关抗原Ki67(Ki67)、天冬氨酸特异性半胱氨酸蛋白酶3(Caspase3)蛋白的表达。starBase在线工具预测结合双荧光素酶报告实验分析LINC00308对miR-634的靶向调控。在PC-3M和DU145细胞中转染miR-634 mimic,利用上述方法检测miR-634在细胞增殖凋亡中的作用。结果前列腺癌组织中的LINC00308表达水平明显高于癌旁组织(P<0.05)。干扰LINC00308显著提高PC-3M和DU145细胞的miR-634的表达水平、G0-G1期细胞比例、细胞凋亡率和Caspase3蛋白水平,明显降低S期细胞比例、Ki67蛋白水平(P<0.05)。LINC00308靶向调控miR-634的表达。转染miR-634 mimic显著增加PC-3M和DU145细胞的G0-G1期细胞比例、凋亡率和Caspase3蛋白水平,明显减少S期细胞比例和Ki67蛋白水平(P<0.05)。结论干扰LINC00308可以促进miR-634的表达,阻滞前列腺癌细胞周期,并诱导细胞凋亡。Objective To investigate whether interfering long-chain non-coding RNA(lncRNA)LINC00308 affects the proliferation and apoptosis of prostate cancer cells by promoting microRNA(miRNA)-634.Methods Real time fluorescent quantitative PCR(QRT PCR)was used to detect the expression of linc00308 in prostate cancer.Prostate cancer cells PC 3M and DU145 were transfected with linc00308 small interfering RNA(Si linc00308).The expression levels of linc00308 and Mir 634 were detected by QRT PCR.The cell cycle and apoptosis were evaluated by flow cytometry.The expression of nuclear associated antigen Ki67(Ki67)and aspartate specific cysteine protease 3(Caspase3)was detected by flow cytometry.Starbase online tool prediction combined with dual luciferase reporter assay was used to analyze the targeting regulation of Mir 634 by linc00308.miR 634 mimic was transfected into PC 3M and DU145 cells,and the effect of miR 634 on cell proliferation and apoptosis was detected by the above method.ResultsThe expression level of LINC00308 in prostate cancer tissue was significantly higher than that in adjacent tissues(P<0.05).Interfering with LINC00308 obviously increased the expression level of miR-634,the proportion of G0-G1 phase cells,the apoptosis rate and the level of Caspase3 protein in PC-3M and DU145 cells,and dramatically reduced the proportion of S phase cells and Ki67 protein level(P<0.05).LINC00308 targets and regulates the expression of miR-634.Transfection of miR-634 mimic remarkably increased the proportion of G0-G1 phase cells,apoptosis rate and Caspase3 protein levels in PC-3M and DU145 cells,and distinctly reduced the proportion of S phase cells and Ki67 protein level(P<0.05).Conclusion Interfering with LINC00308 can promote the expression of miR-634,block the prostate cancer cell cycle,and induce apoptosis.

关 键 词：前列腺癌 LINC00308 miR-634 小干扰RNA 增殖 凋亡 

分 类 号：R737.25[医药卫生—肿瘤]