肝癌细胞PI3K/Akt信号通路在干扰素2α上调ISG15表达中的作用  

The role of PI3K/ Akt signaling pathway in upregulation of ISG15 induced by interferon 2α in hepatocellular carcinoma cells

作　　者：汪蔷华[1] 庞青 王学故 吴华[2] 陈慧娟[1] 赵文俊 李祥 WANG Qiang-hua;PANG Qing;WANG Xue-gu;WU Hua;CHEN Hui-juan;ZHAO Wen-jun;LI Xiang(Department of Infectious Diseases,First Affiliated Hospital of Bengbu Medical College,Bengbu,Anhui 233004,China;不详)

机构地区：[1]蚌埠医学院第一附属医院感染科实验室,安徽蚌埠233004 [2]蚌埠医学院第一附属医院肝胆外科 [3]蚌埠医学院第一附属医院生殖医学中心 [4]蚌埠医学院第一附属医院急诊内科

出　　处：《中华全科医学》2021年第2期182-185,269,共5页Chinese Journal of General Practice

基　　金：安徽省自然科学基金杰出青年项目(2008085J37);蚌埠医学院自然科学基金面上项目(BYKY18111)。

摘　　要：目的探讨肝癌细胞中干扰素2α(IFN-2α)通过磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路上调ISG15表达的机制。方法向培养的肝癌细胞HepG2细胞系中加入IFN-2α和/或PI3K抑制剂(LY294002)。采用蛋白免疫印迹检测各组Akt、磷酸化-蛋白激酶B(p-Akt)和干扰素刺激基因15(ISG15)的蛋白表达水平;采用免疫荧光分析p-Akt和ISG15表达情况。多组间比较采用单因素方差分析。结果 Western印迹分析表明,IFN-2α组中的ISG15表达水平较对照组明显增加,IFN-2α+LY294002组中的ISG15表达水平较IFN-2α组明显降低;免疫染色分析显示,IFN-2α组ISG15荧光强度显著高于对照组和IFN-2α+LY294002组。Western印迹分析显示,和对照组比较,IFN-2α组的p-Akt(Ser473)水平显著降低,而p-Akt(Thr308)水平明显增加;免疫荧光结果显示IFN-2α组中p-Akt(Thr308)的荧光强度显著高于对照组和IFN-2α+LY294002组。结论在HepG2细胞中,IFN-2α通过增加PI3K/Akt通路中的p-Akt(Thr308)上调ISG15的表达。Objective To investigate the mechanism of protein expression of interferon-stimulated gene 15(ISG15) induced by interferon 2α(IFN-2α) via phosphatidy linositol 3-kinase(PI3 K)/protein kinase B(Akt) in hepatocellular carcinoma cells. Methods HepG2 cells were cultured in the media containing IFN-2α and/or PI3 K inhibitor LY294002, and four groups were created: control, IFN-2α, LY294002, IFN-2α+LY294002. The protein expressions of Akt, p-Akt and ISG15 were determined using western blot. Immunofluorescence was used to determine the co-expression of p-Akt and ISG15. Data were analyzed with one-way ANOVA followed by Bonferroni post hoc tests. Results Western blot analysis showed that the protein expression of ISG 15 was significantly increased in the IFN-2α group compared with control and IFN-2α+LY294002 group. Immunostaining analysis showed that the fluorescence intensity of ISG15 in the IFN-2α group was significantly higher than that in control and IFN-2α+LY294002 group. Western blot analysis showed that, in the IFN-2α group, the level of p-Akt(Thr308) was significantly increased, while the p-Akt(Ser473) was significantly decreased, compared with control group. Immunofluorescence results showed that the fluorescence intensity of p-Akt(Thr308) in the IFN-2α group was significantly higher than that in the control group and the IFN-2α+LY294002 group. Conclusion In HepG2 cells, IFN-2α induces the protein expression of ISG 15 via increasing p-Akt(Thr308) in the PI3 K/Akt pathway.

关 键 词：干扰素2α PI3K/AKT信号通路 肝癌细胞 干扰素刺激基因15 

分 类 号：R735.7[医药卫生—肿瘤] R73-36[医药卫生—临床医学]