论文部分内容阅读
目的:探讨PI3K抑制剂LY294002对肺肿瘤细胞增殖影响及其分子机制。方法:培养肺癌细胞系LK2和H460,LY294002阻断PI3K信号通路后,MTT比色法测定细胞生长曲线;FCM分析细胞周期变化;实时RT-PCR、蛋白印迹法分别观察PI3K信号下游蛋白SKP2、p27mRNA及蛋白表达变化。结果:与对照组相比,处理组细胞生长速率受抑制,细胞周期阻滞于G1期。实时RT-PCR及蛋白印迹结果显示,LY294002处理后SKP2mRNA、蛋白表达水平均降低,2种细胞中基因降低量分别为42%和41%,蛋白降低量为65%和55%,p27mRNA表达水平增加了49%和36%,但蛋白表达水平不变。结论:肺肿瘤细胞LK2和H460中PI3K通路通过SKP2,但并不经由p27调节细胞周期,调控细胞增殖。
Objective: To investigate the effect of PI3K inhibitor LY294002 on the proliferation of lung tumor cells and its molecular mechanism. Methods: Lung cancer cell lines LK2 and H460 were cultured, LY294002 blocked the PI3K signaling pathway, cell growth curve was determined by MTT colorimetric assay, cell cycle changes were analyzed by FCM, and PI3K signal downstream proteins SKP2 and p27 mRNA were detected by real-time RT-PCR and Western blot respectively And protein expression changes. Results: Compared with the control group, the growth rate of the treated group was inhibited and the cell cycle was arrested in the G1 phase. Real-time RT-PCR and Western blotting showed that the expression of SKP2mRNA and protein decreased after treatment with LY294002, the gene reduction was 42% and 41%, the protein reduction was 65% and 55%, the expression of p27mRNA was increased 49% and 36%, but the protein expression level unchanged. CONCLUSION: The PI3K pathway in LK2 and H460 lung cancer cells passes through SKP2 but not through p27, which regulates cell cycle and regulates cell proliferation.