阿伐他汀对白血病细胞HL-60增殖和凋亡的影响及其信号通路机制

摘要

Objective:To investigate the effect of atorvastatin on proliferation and apoptosis of leukemia cell line HL-60 and its mechanism of signal pathway.Methods:The leukemia HL-60 cells in logarithmic growth phase were seeded in 96 well plates and were treated with 1,5 and 10 mol/L atorvastatin,then were cultured in the incubator (at 37 ℃,5％ CO2) for 12 h,24 h,48 h.MTT colorimetric method was used to detect the proliferation leukemia cells,the apoptosis of leukemia cells was detected by flow cytometry;the expresion levels of phosphatidylinositol 3-kinase (PI3K),serine threonine protein kinase(ATK) and mTOR at mRNA and protein levels were detected by RT-PCR and Western blot respectively.The experiments included blank control group,the negative control group and drug-treated group.Results:Atorvastatin could inhibit the proliferation of HL-60 cells.The treatment of HL-60 cells with 10 mol/L atorvastatin for 48 hours showed the strongest inhibition rate (39.78 ± 3.00) ％ which was statistically significant different from negative control group (t =4.015,P ＜0.05) and the strongest induction-apoptosis effect on HL-60 cells (43.30±3.92)％,that was statistically significantly different from negative control group (t =3.624,P ＜ 0.05).After treatment with atorvastatin for 48 hours,the expression levels of PI3 K,ATK and mTOR were decreased,in which the effect of 10 mol/L atorvastatin was the most obvious;The expression levels of PI3K,ATK and mTOR were decreased by (37.04 ± 4.15) ％,(53.81 ± 3.25) ％ and (40.62 ± 2.41) respectively,significantly different from the negative control (t =4.806,3.800,4.313,P ＜ 0.05).Conclusion:Atorvastatin may inhibit the proliferation of HL-60 cells and induce apoptosis by inhibiting the PI3K/ATK/mTOR signaling pathway.%目的:探讨阿伐他汀对白血病细胞HL-60增殖和凋亡的影响及信号通路机制.方法:将培养获得的对数生长期的白血病细胞HL-60接种在96孔板,分别给予1、5和10 μmol/L浓度的阿伐他汀,然后放入培养箱中(37℃,5％ CO2)培养(12、24和48 h),用MTT比色法检测白血病细胞的增殖能力.用流式细胞术检测白血病细胞的凋亡变化;RT-PCR法检测PI3K、ATK、mTOR基因mRNA表达水平;Western blot法检测PI3K、ATK、mTOR蛋白表达水平.实验设置空白对照组和阴性对照组.结果:阿伐他汀能够抑制HL-60细胞的增殖,浓度为10 μmol/L的阿伐他汀作用48 h后对HL-60细胞的增殖抑制作用最强,其抑制率为(39.78±3.00)％,与阴性对照组比较,其差异有统计学意义(t =4.015,P＜0.05);对HL-60细胞凋亡的诱导作用最强,其凋亡率为(43.30±3.92)％,与阴性对照组比较,其差异有统计学意义(t =3.624,P＜0.05).同时,阿伐他汀作用48 h后PI3K、ATK、mTOR基因表达水平均有所下降,其中10 μmol/L浓度的阿伐他汀作用最为明显,分别下降了(37.04±4.15)％、(53.81±3.25)％和(40.62±2.41)％.与阴性对照组比较,其差异有统计学意义(t=4.806、3.800、4.313,P＜0.05).结论:阿伐他汀可能通过抑制PI3K/A TK/mTOR信号通路,实现对HL-60细胞增殖的抑制并且诱导其凋亡.