摘要
目的:探讨糖原合成酶激酶-3β(Glycogen synthase kinase-3β,GSK-3β)对肺癌A549细胞增殖的影响及可能机制。方法:MTT法检测不同浓度曲古抑菌素A(TSA)和联合GSK-3β抑制剂SB216763对A549细胞增殖的影响;Western Blot法检测各组细胞中GSK-3β和磷酸化GSK-3β(pGSK-3β)蛋白表达的变化;流式细胞仪分析细胞周期;免疫细胞化学法检测p27蛋白水平。结果:TSA以剂量依赖性抑制A549细胞生长,使细胞周期阻滞于G0/G1期,用SB216763预处理后明显减弱了TSA对A549细胞的生长抑制作用和细胞周期阻滞。TSA对细胞GSK-3β蛋白表达无明显影响,却降低了pGSK-3β蛋白水平,p27蛋白表达增加;抑制剂SB216763使pGSK-3β表达增加,下调p27蛋白水平。结论:GSK-3β参与并促进了TSA对肺癌细胞的生长抑制和细胞周期阻滞效应,其机制可能与GSK-3β对p27蛋白的调节有关。
Objective：To explore the effect of Glycogen synthase kinase-3β （GSK-3 β ）on cell proliferation in human lung adenocarcinoma cell line A549 and its potential mechanisms. Methods： MTT assay was employed to evaluate the effects of trichostatin A （TSA） and pretreatment with SB216763 （a specific inhibitor of GSK-3 β ） on the proliferation of A549 cells. Western Blot was employed to analyze the protein levels of GSK-3β and phosphorylated GSK-3 β （pGSK-3 β ）. Ceil life cycle was examined by flow cytometry analysis. The immunocytochemical method was employed to analyze the protein levels of p27. Results： TSA could inhibit the proliferation of A549 cells in a dose-dependent manner and the cell cycle was arrested in G0/G1 phase in A549 cells treated with TSA. While pre-treatment of A549 cells with SB216763 attenuated TSA induced growth inhibiting and cell cycle arrest. TSA did not alter the levels of GSK-3β but decreased pGSK-3β expression and up-regulated p27 expression. As expected,SB216763 increased the levels of pGSK-3β , consistent with inhibition of GSK-3 β activity. Inhibition of GSK-3 β activity down regulated p27 protein level. Conclusion： GSK-3 β was involved in the inhibition of effect induced by TSA on lung carcinoma cells proliferation process. GSK-3 β may exert this effect by stabilization of p27 which may be one of the downstream molecules.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2009年第2期174-177,共4页
Journal of Chongqing Medical University
