摘要
目的 观察miR-410-3p对前列腺癌细胞PC-3和DU-145增殖和凋亡的影响,并探讨其可能的作用机制.方法 转染miR-NC(对照组)或转染miR-410-3p(实验组)至前列腺癌细胞株PC-3和DU-145;细胞计数试剂盒(CCK-8法)和克隆形成实验检测miR-410-3p对前列腺癌细胞增殖的影响;流式细胞术检测miR-410-3p对细胞凋亡的影响.microRNA.org靶基因预测软件预测miR-410-3p的靶基因;荧光实时定量聚合酶链反应(fluorescence quantitative polymerase chain reaction,qPCR)和免疫印迹法(Western blot)检测血管内皮细胞生长因子C(vascular endothelial growth factor,VEGFC)在mRNA水平和蛋白水平的变化.免疫印迹法检测p-Raf-1和p-ERK1/2蛋白表达.结果 转染miR-410-3p后,细胞的增殖能力明显降低(P<0.05),细胞形成的克隆数明显减少(P<0.05),细胞凋亡明显增加(P<0.01).VEGFC在mRNA水平和蛋白水平的表达明显下降(P<0.01),p-Raf-1和p-ERK1/2蛋白表达明显降低.结论 miR-410-3p可抑制前列腺癌细胞的增殖和促进前列腺癌细胞的凋亡,其可能作用机制为干扰VEGFC基因的表达,为前列腺癌的分子靶向治疗提供新的思路.
Objective To observe the effect of miR-410-3p on the proliferation and apoptosis of prostate cancer cells PC-3 and DU-145,and to explore its possible mechanism.Methods The miR-NC (control group) or miR-410-3p (experimental group) was transfected to prostate cancer cell lines PC-3 and DU-145.The effect of miR-410-3p on the proliferation of prostate cancer cells was detected by cell counting kit (CCK-8) and clone formation experiment.The effect of miR-410-3p on apoptosis was detected by flow cytometry.The target gene of miR-410-3p was predicted by microRNA.org target gene prediction software.Fluorescence real-time quantitative polymerase chain reaction (qPCR) and Western blot were used to detect the changes of VEGFC at mRNA level and protein level.The expression of p-Raf-1 and p-ERK1/2 protein was detected by Western blot.Results After transfection of miR-410-3p,the cell proliferation ability was significantly decreased(P 〈 0.05),the number of clones formed was significantly decreased(P 〈 0.05)and the apoptosis was significantly increased (P 〈 0.01),the expression of VEGFC at mRNA and protein levels were significantly decreased (P 〈0.01).The expression of p-Raf-1 and p-ERK1/2 protein was significantly decreased.Conclusions miR-410-3p can inhibit the proliferation of prostate cancer cells and promote the apoptosis of prostate cancer cells.The possible mechanism is to interfere with the expression of VEGFC gene,which provides a new idea for molecular targeted therapy of prostate cancer.
作者
赵振伶
邵焕军
郝丽娜
于瑞波
Zhao Zhenling1, Shao Huanjun1, Hao Lina2, et al.(1 Department of Urology, People's Hospital of Bozhou City, Bozhou 236800, China; 2 Department of Urology, the First Affiliated Hospital of Kunming Medical University ,Kuming 650032, China)
出处
《国际泌尿系统杂志》
2018年第5期754-758,共5页
International Journal of Urology and Nephrology