下调CENP-W对人脑胶质瘤U87细胞的影响

Effects of CENP-W down-regulation on human glioma U87 cells

目的:研究下调着丝粒蛋白W(centromere protein W,CENP-W)的表达水平对人脑胶质瘤U87细胞的影响。方法:采用小干扰RNA(small interfering RNA,siRNA)抑制人脑胶质瘤U87细胞CENP-W的表达,采用RT-qPCR和Western blot法评价siRNA的干扰效果,采用MTT法、BrdU实验和平板集落形成实验测定细胞的增殖,Transwell小室检测细胞侵袭力的变化,划痕实验检测细胞迁移能力的变化,流式细胞术检测细胞周期及细胞凋亡的改变。结果:MTT实验、平板集落形成实验和BrdU实验显示转染siRNA的实验组U87细胞的增殖能力、集落形成能力较空白对照组及阴性对照组明显降低;Transwell实验和划痕实验显示实验组细胞的迁移、侵袭能力较空白对照组及阴性对照组明显减弱;流式细胞术分析表明实验组细胞的细胞周期被阻滞在G_0/G_1期;实验组细胞凋亡比例较对照组增多(P<0.05)。结论:下调CENP-W的表达可抑制人脑胶质瘤细胞的增殖、迁移和侵袭能力并促进细胞的凋亡,提示CENP-W有望成为人脑胶质瘤基因治疗的候选靶点。

AIM： To study the effect of centromere protein W （CENP-W） down-regulation on human glioma U87 cells. METHODS： Small interfering RNA （siRNA） was used to inhibit the expression of CENP-W in the U87 cells. The interference effect of siRNA was evaluated by RT-qPCR and Western blot. The proliferation of the cells was analyzed by MTT assay, BrdU staining and colony formation experiment. Transwell chamber assay was used to detect the invasion a-bility of the cells. The cell migration ability was measured by a scratch test. The changes of the cell cycle distribution and apoptosis were analyzed by flow cytometry. RESULTS ： The results of MTT assay, colony formation experiment and BrdU staining showed that the cell proliferation and colony formation abilities in experimental group were significantly lower than those in control group and negative control group. The results of Transwell and scratch experiments showed that the migra-tion and invasion abilities in experimental group were weaker than those in blank control group and negative control group. The results of flow cytometry analysis showed that the cell cycle distribution in experimental group was arrested in G0/G ） phase. The percentage of apoptotic cells in experimental group was higher than that in control group （ P 〈 0. 05 ） . CON-CLUSION Down-regulation of CENP-W expression inhibits the proliferation, migration and invasion of human gliomacells and promotes the apoptosis of the cells, suggesting that CENP-W may be a potential target of gene therapy for human glioma.