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干扰素诱导肝癌细胞ADAR1的表达模式及HBV复制抑制
Effect of interferons on expression of adenosine deaminase acting on RNA 1 and hepatitis B virus replication in human hepatocellular carcinoma cells
in vitro - 第三军医大学学报 2019年41卷第14期 页码：1336-1342
- 作者机构：
1.400016 重庆，重庆医科大学:感染性疾病分子生物学教育部重点实验室
2.400016 重庆，重庆医科大学:基础医学院病原生物学教研室，分子医学与肿瘤研究中心
- 作者简介：
[涂增，E-mail:tuz1980@126.comTU Zeng, E-mail:tuz1980@126.com
- 基金信息：重庆市基础科学与前沿技术研究专项(CSTC2015jcyjA10006);国家自然科学基金青年科学基金(81501751);重庆市博士后科学基金(Xm2014006)
DOI：10.16016/j.1000-5404.201901149
中图分类号： R735.7; R966; R977.6收稿：2019-01-18，
修回：2019-03-10，
纸质出版：2019-07-30
- 稿件说明：
移动端阅览
李怡敏, 袁琳, 杨生永, 等. 干扰素诱导肝癌细胞ADAR1的表达模式及HBV复制抑制[J]. 第三军医大学学报, 2019,41(14):1336-1342.
Yimin LI, Lin YUAN, Shengyong YANG, et al. Effect of interferons on expression of adenosine deaminase acting on RNA 1 and hepatitis B virus replication in human hepatocellular carcinoma cells
in vitro [J]. Journal of Third Military Medical University, 2019, 41(14): 1336-1342.李怡敏, 袁琳, 杨生永, 等. 干扰素诱导肝癌细胞ADAR1的表达模式及HBV复制抑制[J]. 第三军医大学学报, 2019,41(14):1336-1342. DOI： 10.16016/j.1000-5404.201901149.
Yimin LI, Lin YUAN, Shengyong YANG, et al. Effect of interferons on expression of adenosine deaminase acting on RNA 1 and hepatitis B virus replication in human hepatocellular carcinoma cells
in vitro [J]. Journal of Third Military Medical University, 2019, 41(14): 1336-1342. DOI： 10.16016/j.1000-5404.201901149.
摘要
目的
2
探讨Ⅰ型干扰素(IFN-α、IFN-β)和Ⅱ型干扰素(IFN-γ)在肝癌细胞中诱导ADAR1(P110和P150)的表达模式及对HBV复制的作用。
方法
2
不同浓度IFN-α(0、200、500、1 000、2 000 U/mL)、IFN-β(0、200、1 000 U/mL)和IFN-γ(0、200、1 000 U/mL)分别处理肝癌细胞(Huh7和HepG2)，Western blot和RT-qPCR检测肝癌细胞中IFN作用后ADAR1的表达变化；Huh7细胞瞬时转染HBV复制型质粒，IFN-α作用后，Southern blot分析HBV DNA复制中间体水平。
结果
2
Ⅰ型IFN(IFN-α、IFN-β)作用Huh7和HepG2细胞后，ADAR1 P110蛋白水平增加(
P
＜0.05)，同时ADAR1 P150表达水平明显增高(
P
＜0.01)，且ADAR1 P150的表达呈现IFN浓度依赖性；Ⅱ型IFN(IFN-γ)作用Huh7和HepG2细胞后，ADAR1 P110和P150蛋白表达水平没有发生明显变化。IFN-α明显抑制Huh7细胞中HBV复制(
P
＜0.05)，与浓度梯度呈负相关。
结论
2
Ⅰ型IFN可诱导肝癌细胞中ADAR1 P150表达，IFN-α抑制Huh7细胞中HBV复制，其作用可能与ADAR1相关。
Abstract
Objective
2
To investigate the effect of type Ⅰ interferons (IFN-α and IFN-β) and type Ⅱ interferon (IFN-γ) on the expression of adenosine deaminase acting on RNA 1 (ADAR1
P110 and P150) and hepatitis B virus (HBV) replication in hepatocellular carcinoma (HCC) cells.
Methods
2
Western blotting and qRT-PCR were used to detect the expression of ADAR1 in human HCC cell lines HepG2 and Huh7 following treatment with different concentrations of IFN-α
IFN-β or IFN-γ. Southern blotting was employed to detect the changes in HBV DNA level in response to IFN-α treatment in Huh7 and HepG2 cells transfected with HBV replication plasmids.
Results
2
In Huh7 and HepG2 cells
treatments with IFN-α and IFN-β significantly increased the protein expression of ADAR1 P150 in a time- and dose-dependent manner (
P
<
0.01) and also obviously augmented the protein level of ADAR1 P110; IFN-γ did not significantly affected the expression of eithe
r ADAR1 P110 or ADAR1 P150. IFN-α treatment dose-dependently inhibited HBV DNA replication in Huh7 cells (
P
<
0.05).
Conclusion
2
Type Ⅰ interferons time- and dose-dependently increase the expression of ADAR1 in Huh7 and HepG2 cells
suggesting a possible role of ADAR1 in mediating the anti-HBV effect of IFN-α.
关键词
Keywords
references
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