Dai Jing, Liu Tao, Wang Kai, Pang Ying-Xu, Wang Qiong, Chen Zhen-Zhu
Department of Hematology, Zhoukou Municipal Central Hospital, Zhoukou 466000, Henan Province, China.
Department of Hematology, Zhoukou Municipal Central Hospital, Zhoukou 466000, Henan Province, China. E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2017 Apr;25(2):403-407. doi: 10.7534/j.issn.1009-2137.2017.02.017.
To investigate the effect of atorvastatin on proliferation and apoptosis of leukemia cell line HL-60 and its mechanism of signal pathway.
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 °C, 5% CO) 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.
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 PI3K,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).
Atorvastatin may inhibit the proliferation of HL-60 cells and induce apoptosis by inhibiting the PI3K/ATK/mTOR signaling pathway.
探讨阿托伐他汀对白血病细胞株HL-60增殖和凋亡的影响及其信号通路机制。
将对数生长期的白血病HL-60细胞接种于96孔板，分别用1、5和10 μmol/L阿托伐他汀处理，然后在培养箱（37℃，5% CO₂）中培养12 h、24 h、48 h。采用MTT比色法检测白血病细胞的增殖情况，流式细胞术检测白血病细胞的凋亡情况；分别用RT-PCR和Western blot检测磷脂酰肌醇3激酶（PI3K）、丝氨酸苏氨酸蛋白激酶（AKT）和哺乳动物雷帕霉素靶蛋白（mTOR）在mRNA和蛋白水平的表达。实验分为空白对照组、阴性对照组和药物处理组。
阿托伐他汀可抑制HL-60细胞的增殖。10 μmol/L阿托伐他汀处理HL-60细胞48小时的抑制率最强（39.78±3.00）%，与阴性对照组相比差异有统计学意义（t=4.015，P<0.05），对HL-60细胞的诱导凋亡作用最强（43.30±3.92）%，与阴性对照组相比差异有统计学意义（t=3.624，P<0.05）。阿托伐他汀处理48小时后，PI3K、AKT和mTOR的表达水平降低，其中10 μmol/L阿托伐他汀的作用最明显；PI3K、AKT和mTOR的表达水平分别降低（37.04±4.15）%、（53.81±3.25）%和（40.62±2.41）%，与阴性对照组相比差异有统计学意义（t=4.806、3.800、4.313，P<0.05）。
阿托伐他汀可能通过抑制PI3K/AKT/mTOR信号通路抑制HL-60细胞的增殖并诱导其凋亡。
