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目的研究抑制eEF-2激酶对人脑胶质瘤LN229细胞增殖作用的影响,并初步探讨其可能的分子机制。方法采用四甲基偶氮唑盐(MTT)比色法检测抑制eEF-2激酶后LN229细胞的增殖能力,台盼蓝染色法检测细胞的存活能力,相差显微镜观察细胞形态学的变化,流式细胞仪分析细胞周期的分布,Western blot方法检测细胞phospho-eEF2和Cleaved Caspase-3、PARP蛋白表达水平的变化。结果抑制eEF-2激酶对人脑胶质瘤LN229细胞的增殖有显著抑制作用(P<0.05或0.01),呈现时间和剂量依赖性趋势;作用24 h后,随着给药浓度的增加,LN229细胞的活力逐渐降低(P<0.05或0.01),且细胞形态也逐渐发生明显改变;NH125(0.5μmol/L)作用24 h后,LN229在S期阻滞,且出现小凋亡峰,phospho-eEF2(Thr56)的表达明显下降,Cleaved Caspase-3、Cleaved PARP的表达显著增加(P<0.05)。结论抑制eEF-2激酶后,人脑胶质瘤LN229的细胞增殖作用明显被抑制,细胞活力降低,该作用可能与其诱导的细胞周期阻滞以及其诱导的Caspase-3、PARP激活相关细胞凋亡途径有关。
Objective To study the effect of inhibiting eEF-2 kinase on the proliferation of human glioma LN229 cells and to explore its possible molecular mechanism. Methods MTT assay was used to detect the proliferation of LN229 cells after eEF-2 kinase inhibition. Cell viability was assayed by trypan blue staining. Cell morphological changes were observed by phase contrast microscopy. Flow cytometry Cytokines were used to analyze the cell cycle distribution. Western blot was used to detect the expression of phospho-eEF2, Cleaved Caspase-3 and PARP protein. Results The inhibitory effect of eEF-2 kinase on the proliferation of human glioma LN229 cells was significantly inhibited (P <0.05 or 0.01), showing a time-and dose-dependent trend. After 24 h treatment, with the increasing concentration of LN229 The cell viability decreased gradually (P <0.05 or 0.01), and the cell morphology also gradually changed. After treated with NH125 (0.5μmol / L) for 24 h, LN229 blocked in S phase with small apoptotic peak and phospho- The expression of eEF2 (Thr56) was significantly decreased, while the expression of Cleaved Caspase-3 and Cleaved PARP was significantly increased (P <0.05). Conclusion Inhibition of eEF-2 kinase inhibits the proliferation of human glioma LN229 cells and decreases the viability of LN229 cells, which may be related to the cell cycle arrest induced by eEF-2 kinase and the apoptosis of LN229 cells induced by Caspase-3 and PARP Way related.