miR-410-3p对前列腺癌细胞增殖和凋亡的影响及作用机制研究

Effects of miR-410-3p on proliferation and apoptosis of prostate cancer cells and its mechanism

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.