肝癌细胞PI3K/Akt信号通路在干扰素2α上调ISG15表达中的作用

汪蔷华; 庞青; 王学故; 吴华; 陈慧娟; 赵文俊; 李祥

doi:10.16766/j.cnki.issn.1674-4152.001762

肝癌细胞PI3K/Akt信号通路在干扰素2α上调ISG15表达中的作用

doi: 10.16766/j.cnki.issn.1674-4152.001762

汪蔷华¹,
庞青²,
王学故³,
吴华²,
陈慧娟¹,
赵文俊⁴,
李祥^3, ,

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，安徽蚌埠 233004
3.
蚌埠医学院第一附属医院生殖医学中心，安徽蚌埠 233004
4.
蚌埠医学院第一附属医院急诊内科，安徽蚌埠 233004

基金项目:

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

详细信息

通讯作者:
李祥，E-mail：li19850115@163.com

中图分类号: R735.7 R73-36
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- 文章访问数: 120
- HTML全文浏览量: 107
- PDF下载量: 1
- 被引次数: 0
出版历程
- 收稿日期: 2020-10-16
- 网络出版日期: 2022-02-19

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

1.
Department of Infectious Diseases, First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233004, China

摘要

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。
- 干扰素2α /
- PI3K/Akt信号通路 /
- 肝癌细胞 /
- 干扰素刺激基因15
Abstract: 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 (PI3K)/protein kinase B (Akt) in hepatocellular carcinoma cells. Methods HepG2 cells were cultured in the media containing IFN-2α and/or PI3K 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 PI3K/Akt pathway.
- Interferon 2α /
- PI3K/Akt signaling pathway /
- Hepatocellular carcinoma cells /
- Interferon stimulated gene 15

HTML全文

图 1 ISG15在肝癌患者肝癌组织中的表达及生存分析

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图 2 蛋白免疫印迹和免疫荧光分析各组HepG2细胞中ISG15的表达水平

注：A为HepG2中ISG15蛋白表达的Western blotting代表图；B为HepG2中ISG15蛋白表达的Western blotting条带分析；^aP < 0.05；^bP < 0.001。

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图 3 各组HepG2细胞中p-Akt(Ser473)、p-Akt(Thr308)和Akt的表达

注：A为蛋白印迹检测各组HepG2细胞中p-Akt(Ser473)、p-Akt(Thr308)和Akt的Western blotting的代表条带；B为各组p-Akt(Ser473)、p-Akt(Thr308)和Akt灰度值统计；^aP<0.05；^bP < 0.001；^cP < 0.0001。

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图 4 各组HepG2细胞中p-Akt(Thr308)和ISG15的免疫荧光

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