他汀类药物通过抑制Akt通路调控急性T淋巴细胞白血病细胞增殖与凋亡

Statins Regulate the Proliferation and Apoptosis of T-ALL Cells through the Inhibition of Akt Pathway

目的:探讨他汀类药物对急性T淋巴细胞白血病(T-ALL)细胞增殖、凋亡的影响及其可能的作用机制。方法:Jurkat及CCRF-CEM细胞经不同浓度的氟伐他汀和辛伐他汀药物处理24 h,CCK-8法检测细胞增殖抑制情况;Ed U法检测细胞DNA复制情况;Annexin V/7-AAD双染法分析细胞凋亡情况;流式细胞术分析细胞周期的变化;免疫印迹法检测Cyclin D1、p21、p27、BAX、BCL-2及p-Akt蛋白的表达。结果:氟伐他汀、辛伐他汀均可抑制Jurkat及CCRF-CEM细胞增殖,且抑制效应呈一定的剂量依赖性。氟伐他汀浓度为0.2 mmol/L时,Jurkat和CCRF-CEM细胞抑制率分别达41.14%和57.08%;而辛伐他汀浓度为0.2 mmol/L时,Jurkat和CCRF-CEM细胞抑制率分别达68.42%和77.10%,均接近或超过半数抑制,与溶剂对照组比较有显著差异(P<0.05)。不同浓度的氟伐他汀及辛伐他汀分别作用于Jurkat和CCRF-CEM细胞24 h,其早期凋亡和晚期凋亡的细胞比例均升高,且于0.2 mmol/L和0.3 mmol/L药物浓度时,其总凋亡比例显著升高,与溶剂对照组相比有显著差异(P<0.05)。细胞周期检测结果显示,氟伐他汀及辛伐他汀诱导Jurkat和CCRF-CEM细胞G_1期阻滞。Western blot检测结果显示,Jurkat和CCRF-CEM细胞经氟伐他汀或辛伐他汀处理后,BAX、p21和p27表达量逐渐升高,Cyclin D1、BCL-2和p-Akt表达量逐渐减低。结论:他汀类药物可通过抑制Akt信号通路抑制T-ALL细胞的增殖,并诱导其凋亡。

Objective： To investigate the effect of Statins on proliferation and apoptosis in human acute T lymphocytic leukemia （T-ALL） cells and its possible mechanism. Methods： Jurkat and CCRF - CEM cells were cultured in different concentrations of Fluvastatin and Simvastatin for 24 h respectively. Then, the cell growth inhibition level was defected by CCK-8; the DNA replication was analyzed by EdU; the cell apoptosis was analyzed by Annexin V/7-AAD double labeling; the cell cycle changes were analyzed by flow cytometry; the expressions of Cyclin D1, p21, p27, BAX, BCL-2 and p-Akt were determined by Western blot. Results： Fluvastatin and Sirnvastatin both significantly inhibited the growth of Jurkat and CCRF - CEM cells in a dose-dependent manner. The inhibitory rate of Jurkat and CCRF- CEM cells at 0. 2 mmol/L Fluvastatin was 41.14% and 57. 08% respectively, while the 0. 2 mmol/L Simvastatin could supress 68.42% of Jurkat and 77.10% of CCRF - CEM cells. Half or more than half of cell inhibition were observed in Statins - treated groups with significantly statistical differences, compared with the control groups （ P 〈 0. 05）. After the Jurkat and CCRF - CEM ceils were treated with Fluvastation and Simvastation of different concentrations for 24 hours, the proportion of early and later apoptotic cells both increased; moreover, the total apoptotic rate increased significantly （ P 〈 0. 05 ） at 0.2 mmol/L and 0.3 mmol/L concentration of Fluvastatin and Simvastatin. The detection of cell cycle showed that both of Jurkat and CCRF - CEM cells were arrested in G1 phase. Western blot revealed that, in comparison with the control group, the expressions of BAX, p21 and p27 in cells treated with Statins were up-regulated, while Cyclin D1, BCL-2 and p-Akt expressions were down-regulated. Conclusion： Statins can suppress T-ALL cell proliferation and induce cell apoptosis through the inhibition of Akt pathway.