目的探讨节律基因Clock对胶质瘤C6细胞增殖的影响及其机制。方法通过小片段干扰RNA(small interfering,siRNA)特异性沉默C6细胞Clock基因的表达。采用CCK-8试剂盒检测C6细胞的增殖情况,并利用流式细胞仪检测Clock基因沉默后C6细胞的细胞周期分布,通过荧光定量Q-PCR检测β-catenin和Tcf1mRNA的表达水平。结果与空白对照组和阴性对照组相比,SiClock转染组C6细胞增殖活力明显降低,S期细胞所占比例明显下降,同时β-catenin和Tcf1mRNA的表达也明显降低(均P〈0.05)。结论干扰节律基因Clock表达,能够抑制胶质瘤细胞的生长。其机制可能是通过Wnt/β-catenin信号通路调节其作用。
Objective To investigate the effect of circadian gene Clock on glioma C6 cells and its mechanism.Methods The expression level of Clock gene in C6 cells were specifically knocked down by RNA interference.The effect of C6 cells on cell growth and proliferation was demonstrated with CCK-8assay.The cell distribution in cell cycle was detected by flow cytometry.The expression of β-catenin and Tcf1 were detected by Q-PCR.Results Compared with the blank control group and the negative control group,the cells transfected with SiClock had the lower rate of cellular proliferative capacity.Meanwhile,the proportion of S phase cells decreased obviously.The mRNA expression of β-catenin and Tcf1 were significantly reduced.Conclusion Clock gene knocked down inhibited the proliferation of glioma cell,which mechanism might involve the Wnt/β-catenin signaling pathway.