摘要
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 investigate the effect of eEF-2 kinase inhibitor on proliferation of glioma cell line LN229.Method After eEF-2 kinase was inhibited,proliferation and viability of glioma LN229 cell was determined by MTT and trypan blue assay respectively.Morphological changes of tumor cells were detected by light microscope.Cell cycle was analyzed by flow cytometry.The levels of phospho-eEF2,Cleaved Casepase-3 and PARP proteins were measured by Western blot analysis.Results Following the inhibition of eEF-2 kinase,the proliferation of LN229 giloma cells was significantly inhibited in a time-concentration dependent manner（P0.05 or 0.01）.Twenty-four hours after treatment,along with the increasing concentrations of NH125,LN229 cell viability significantly decreased（P0.05 or 0.01）,and cell morphology altered.After being treated with NH125（0.5 μmol/L）for 24 h,LN229 giloma cell cycle was arrested at S phase and with a small apoptotic peak,the expression of phospho-eEF2（Thr56） was decreased,and the expressions of Cleaved Caspase-3,Cleaved PARP were increased.Conclusion Inhibition of eEF-2 kinase caused a significant suppression of proliferation and a significant reduction of the cell viability of human glioma LN229 cells.The underlying mechanisms may be related to induction of cell cycle arrest and the activation of Casepase-3 and PARP-related apoptosis pathway.
出处
《苏州大学学报（医学版）》
CAS
2012年第5期623-628,共6页
Suzhou University Journal of Medical Science
基金
江苏省自然科学基金资助项目(BK20102242)
江苏省高校自然科学基础研究面上项目(08KJB310010)
