血管紧张素Ⅱ对白血病HL-60细胞株骨桥蛋白表达的影响及机制

The Effect and Mechanism on Expression of Osteopontin in Leukemic Cell Line HL-60 Induced by Antagonism Ⅱ

目的:研究血管紧张素Ⅱ(AngⅡ)对白血病HL-60骨桥蛋白表达的影响及机制。方法:体外培养HL-60细胞株,予Ang II(终浓度10-9、10-8、10-7、10-6 mol/L)刺激24 h或予AngⅡ(终浓度10-7 mol/L)刺激12h、24 h、48 h;先经PI3K/AKT抑制剂LY294002(40 umol/L)预处理1 h,再予10-7 mol/L Ang II刺激24 h,收集细胞,采用western-blot法检测OPN表达情况。结果:AngⅡ促进HL-60细胞表达OPN,在一定浓度及时间内,其表达量随着AngⅡ浓度和时间的增加而增加,呈剂量和时间依赖性关系;经LY294002预处理再予AngⅡ刺激的HL-60细胞,与对照组比较,OPN表达明显抑制(P<0.05)。结论 :AngⅡ经PI3K/AKT信号通路诱导OPN的表达。

Objective： To investigate the effect and mechanism of antagonism Ⅱ（Ang Ⅱ ） on expression of osteopontin（OPN）in leukemic cell line HL-60. Methods： Leukemic cell line HL-60 was cultured in RPMI 1640 and stimulated by Ang Ⅱ at the final concentration of 10^-9, 10^-8, 10^-7, 10^-6 mol/L for 24 h or at final concentration of 10^-7 mol/L for 12 h,24 h and 4 8h. HL-60 cells were pretreated with LY294002,a specific inhibitor of PI3K/AKT signaling pathway, at final concentration of 40 umol/L for lh ,followed by incubation with Ang Ⅱ at final concentration of 10^-7mol/L for 24h.The cells were collected. The expression of OPN was detected by Western blotting. Results：In HL-60 ceils ,the expression of OPN was apparently elevated when incubated with Ang I/.With the increase in Ang Ⅱ concentration or the extension of incubation hours, the expression of OPN was increasd gradually in a dose- and time- dependent manner. HL-60 cells pretreated with LY294002 and incubated with had a significant decrease in the OPN expression as compared with control group （P〈O.05）. Conchlusion：The expression of OPN in Ang Ⅱ-induced HL-60 cells is regulated by the PI3K/AKT signaling pathway.