siRNA沉默CCN5抑制人胶质母细胞瘤细胞的生长

目的研究小干扰RNA(siRNA)沉默高半胱氨酸蛋白61/结缔组织生长因子/肾母细胞瘤过度表达基因5(CCN5)对人胶质母细胞瘤细胞系生长的影响。方法 Western blot方法检测胶质母细胞瘤患者脑组织CNN5蛋白水平的表达;合成CCN5 siRNA小片段,并,筛选最佳干扰片段;siRNA沉默CCN5在胶质母细胞瘤细胞系中的表达,噻唑兰(MTT)方法检测细胞的生长;CCN5沉默表达后,检测半胱氨酸的天冬氨酸蛋白水解酶3(caspase-3)的酶活性。结果胶质母细胞瘤患者脑组织中CCN5蛋白表达水平较正常人脑组织中高;siRNA沉默CCN5表达抑制细胞的生长并促进凋亡效应基因caspase-3活性升高。结论 CCN5基因沉默抑制人胶质母细胞瘤细胞系的生长,为探讨CCN5在胶质母细胞瘤细胞中的作用奠定基础,并为胶质母细胞瘤的基因治疗提供潜在的靶标基因。

CCN5 silenced by siRNA inhibits the growth of human glioblastoma cells

Objective The effect of connective tissue growth factor/cysteine-rich 61/nephroblastoma overexpressed 5 （CCN5） small interference RNA （siR NA） on the growth of human glioma cells is investigated.Methods CCN5 protein levels in brain tissues of glioblastoma patients were detected by Western blot.The synthesized CCN5-siRNA was ttransfected into cells by liposome and the transfection efficiency was confirmed by Western blot.The effect of cell growth was analyzed by 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay.Meanwhile,the activity of caspase-3 was evaluated by caspase-3 activity assay kit.Results The CCN5 protein expression in glioblastoma patients was higher than that of normal persons.Compared with control group,CCN5-siRNA significantly inhibited the growth of glioblastoma cells and promoted the activity of cysteinyl aspartate specific proteinase 3 （caspase-3）.Conclusion This study demonstrates that CCN5 sileenced by siRNA significantly suppresses the growth of glioblastoma cells which indicates that CCN5 plays a critical role in the gliomagenesis and development of grioma and may be a potential target for human glioma therapy.