敲减CDKL5对人胶质母细胞瘤细胞生长和细胞周期的影响

Effect of CDKL5 knockdown on growth and cell cycle of human glioblastoma cells

目的探讨敲减周期蛋白依赖型激酶样5(cyclin-dependent kinase-like 5,CDKL5)对人胶质母细胞瘤细胞生长和细胞周期的影响,为胶质母细胞瘤的治疗提供新思路。方法本研究使用人胶质母细胞瘤细胞U87,使用慢病毒感染的方法敲减CDKL5基因。利用Western blot显示CDKL5的细胞定位以及特异性的小发卡RNA敲减效率。使用血细胞计数法绘制4 d U87的细胞生长曲线,同时利用MTT检测细胞活性。使用流式细胞仪技术检测细胞周期。结果在U87人胶质母细胞瘤细胞中,CDKL5在细胞核和细胞质当中都有表达,我们发现使用shRNA敲减CDKL5后,胞质中的CDKL5显著减少,细胞形态也发生明显改变。敲减CDKL5同时抑制人胶质母细胞瘤细胞的体外增,引起细胞周期停滞。结论敲减CDKL5可以显著抑制人胶质母细胞瘤细胞增殖,引起细胞周期停滞,为人胶质母细胞瘤的治疗提供了一个新的理论方法。

Objective To investigate the effects of CDKL5 knockdown on the cell growth and cycle of human glioblastoma cells,and to provide new strategy for the treatment of glioblastoma.Methods In this study,CDKL5 gene was knock down the using lentivirus infection in human glioblastoma U87 cells.Western blot analyais confirmed CDKL5 cell localization and specific small hairpin RNA knockdown efficiency.Hematocyte counting method was used to draw the cell growth curve of U87 for 4 days,and MTT was used to detect cell activity.Cell cycle was detected using flow cytometry.Results CDKL5 is expressed in both the nucleus and the cytoplasm in U87 human glioblastoma cells.Our results showed that CDKL5 was significantly reduced in the cytoplasm after shRNA knockdown using shRNA,and the cell morphology also changed significantly.Knockdown CDKL5 also inhibited the growth of human glioblastoma cells in vitro,causing cell cycle arrest.Conclusion CDKL5 knockdown can significantly inhibit human glioblastoma cell proliferation and cause cell cycle arrest,providing a new theoretical approach for the treatment of human glioblastoma.