靶向沉默甲基转移酶3对前列腺癌细胞增殖、侵袭和克隆能力的影响

Effect of silencing METTL3 on the proliferation,invasion and cloning of prostate cancer cells

目的探讨小干扰RNA靶向沉默甲基转移酶3(METTL3)基因对前列腺癌DU145、PC-3细胞增殖、侵袭、克隆能力的影响。方法将DU145、PC-3细胞随机设立空白对照组(无任何处理)、Mock组(仅加转染试剂)、阴性对照组(阴性siRNA干扰片段)、干扰组(METTL3-siRNA干扰片段),运用Lipofectamine^TM 2000脂质体转染DU145、PC-3细胞,使用qRT-PCR,Western Blot实验分别检测METTL3的mRNA和蛋白表达情况,采用CCK-8、Transwell、平板克隆实验分别检测各组细胞增殖、侵袭、克隆能力。结果DU145、PC-3细胞干扰组转染siRNA-1400干扰片段48 h后,METTL3 mRNA的干扰效率分别为90%、72%(P<0.01),METTL3蛋白的干扰效率分别为87.2%、97.8%(P<0.01)。与空白对照组和阴性对照组比较,DU145、PC-3细胞干扰组细胞增殖、侵袭、克隆形成能力均显著下降(均P<0.05)。结论METTL3基因下调后将明显抑制前列腺癌细胞的增殖、侵袭、克隆能力,可能是前列腺癌治疗的一个新型分子靶点。

Objective To investigate the effect of small interfering RNA targeted silencing METTL3 gene on proliferation,invasion and cloning of prostate cancer DU145 and PC-3 cells.Methods DU145 and PC-3 cells were randomly divided into blank control group(without any treatment),mock group(only with transfection reagent),negative control group(negative interference fragment),interference group(METTL3-siRNA interference fragment).Lipofectamine^TM 2000 was used to transfect DU145 and PC-3 cells.mRNA and protein expression of METTL3 were detected by qRT-PCR and Western Blot respectively.The proliferation,invasion and clone ability of cells in each group were detected by CCK-8,transwell and plate clone test respectively.Results When the interference group transfected siRNA-1400 for 48 h,the interference efficiencies of METTL3 mRNA in DU145 and PC-3 cells were 90%and 72%(P<0.01).The interference efficiencies of METTL3 protein in DU145 and PC-3 cells were 87.2%and 97.8%,respectively(P<0.01).Compared with the blank control group and the negative control group,the proliferation,invasion and colony forming ability of the interference group in DU145 and PC-3 cells were decreased significantly(P<0.05).Conclusion Downregulation of METTL3 gene will significantly inhibits the proliferation,invasion and cloning of prostate cancer cells,and may be a new molecular target in prostate cancer therapy.