目的探讨氯化锂(lithium chloride,LiCl)对人肺泡Ⅱ型上皮细胞来源的A549细胞在细胞增殖和细胞周期方面的影响。方法以不同浓度的LiCl作用A549细胞24h后,采用Cell Counting Kit-8(CCK-8)检测各组细胞的增殖活性;利用流式细胞术进行细胞周期分析;免疫细胞荧光技术检测糖原合成酶激酶3β(glycogen synthase kinase3β,GSK3β)在各组细胞中的表达;并通过Western blot检测LiCl对GSK3β、磷酸化GSK3β(p—GSK3β)及CyclinD1蛋白质表达水平的影响。结果LiCl能提高A549细胞的增殖活性。且随着LiCl浓度的升高,处于G1期的细胞数量减少,而S期细胞数量增多,与正常对照组相比,差异具统计学意义(10mmol/L组:P〈0.05;20mmol/L组:P〈0.01)。LiCl能使GSK3β表达降低而无酶活性的p-GSK3β表达升高;同时上调CyclinD1的表达水平。结论LiCl能加快A549细胞的G1/S期转换并促进增殖,GSK3β的活性受抑从而减少Cyclin D1的降解是其可能的机制。
Objective To investigate the effect of lithium chloride (LiCl) on cell proliferation and cell cycle progression in alveolar epithelial type Ⅱ cell-derived A549 cells. Methods After A549 cells were treated with different concentrations of LiCl for 24 h, cell viability of each group was measured by cell counting Kit-8, and cell cycle was analyzed by flow cytometry. The immunofluorescence was studied and the expression of glycogen synthase kinase 3β (GSK3β), phosphorylated GSK3β (p- GSK3β) and Cyclin D1 were detected by Western blotting. Results LiCl could elevate cell viability of A549. Moreover, with the increase of LiCl concentrations, percentage of cells in G1 phase was decreased, and that in S phase increased. There was significant difference between control group and LiCl-treated groups (10 mmol/L group, P〈0.05;20 mmol/L group, P〈0. 191). Meanwhile, LiCl treatment could decrease the expression level of GSK3β and up-regulate that of kinase-dead p-GSK3β as well as Cyclin D1. Conclusion LiCl promoted G1/S transition and cell proliferation in A549 cells possibly by inhibition of GSK3β activity and consequent stabilization of Cyclin D1.