中文English
ISSN 1001-5256 (Print)
ISSN 2097-3497 (Online)
CN 22-1108/R

Effect of hepatitis B x gene on the expression of major histocompatibility complex class I chain-related gene A-A5. 1,invasion,and migration of HepG2. 2. 15 cells

DOI: 10.3969/j.issn.1001-5256.2020.04.020
Research funding:

 

  • Received Date: 2019-10-24
  • Published Date: 2020-04-20
  • Objective To investigate the effect of hepatitis B x( HBx) gene on the expression of major histocompatibility complex class I chain-related gene A( MICA)-A5. 1,invasion,and migration of HepG2. 2. 15 cells. Methods HepG2. 2. 15 cells( HepG2 cells with the insertion and continuous expression of whole HBV genome) were cultured in vitro and were randomly divided into control group,HBx overexpression plasmid group,HBx empty plasmid group,HBx siRNA group,and HBx siRNA negative control group. After the transfection of plasmid or siRNA,CCK-8 assay was used to measure cell proliferation after 24 and 48 hours to screen out the appropriate duration of drug action; the Transwell invasion test and wound-healing assay were used to observe the changes in cell invasion and migration abilities;immunoblotting was used to measure the expression of HBx,MICA-A5. 1,and the marker proteins for epithelial-mesenchymal transition E-cadherin and vimentin; ELISA was used to measure the level of soluble MICA( s MICA) in cell culture medium. A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the SNK q test was used for further comparison between two groups. Results Compared with the control group after 24 hours,the HBx overexpression plasmid group had a significant increase in cell viability and the HBx siRNA group had a significant reduction in cell viability( q = 8. 268 and 4. 365,P < 0. 001 and 0. 036); compared with the control group after 48 hours,the HBx overexpression plasmid group had a significant increase in cell viability and the HBx siRNA group had a significant reduction in cell viability( q = 12. 680 and 7. 523,both P < 0. 001). Compared with the control group,the HBx overexpression plasmid group had significant increases in the protein expression of HBx,MICA,and vimentin in cells,the level of s MICA in cell culture medium,the number of migrating cells,and the number of cells out of the Transwell chamber,as well as a significant reduction in the expression of E-cadherin( q = 9. 427,6. 697,10. 500,5. 042,22. 740,15. 720,and 5. 258,all P < 0. 05); the HBx siRNA group had significant reductions in the protein expression of HBx,MICA,and vimentin in cells,the level of s MICA in cell culture medium,the number of migrating cells,and the number of cells out of the Transwell chamber,as well as a significant increase in the expression of E-cadherin( q = 8. 133,8. 828,7. 616,7. 673,5. 391,7. 694,and 6. 226,all P < 0. 05). Conclusion The HBx gene regulates theexpression of MICA-A5. 1 in HepG2. 2. 15 cells and the invasion and migration of HepG2. 2. 15 cells,and upregulation of the HBx gene can promote the expression of MICA-A5. 1 and enhance the invasion and migration abilities of HepG2. 2. 15 cells.

     

  • [1] SHIRATA C,HASEGAWA K,KOKUDO T,et al. Liver resection for hepatocellular carcinoma in patients with renal dysfunction[J]. World J Surg,2018,42(12):4054-4062.
    [2] ABOU-ALFA GK,MEYER T,CHENG AL,et al. Cabozantinib in patients with advanced and progressing hepatocellular carcinoma[J]. N Engl J Med,2018,379(1):54-63.
    [3] GARDINI AC,FOSCHI FG,CONTI F,et al. Immune inflammation indicators and ALBI score to predict occurrence and recurrence of hepatocellular carcinoma in HCV-related cirrhosis treated with direct-acting antivirals[J]. Dig Liver Dis,2018,50(1):50-51.
    [4] WANG M,XI D,NING Q. Virus-induced hepatocellular carcinoma with special emphasis on HBV[J]. Hepatol Int,2017,11(2):171-180.
    [5] GAO Q,WANG K,CHEN K,et al. HBx protein-mediated ATOH1 downregulation suppresses ARID2 expression and promotes hepatocellular carcinoma.[J]. Cancer Sci,2017,108(7):1328-1337.
    [6] SHI T,HUA Q,MA Z,et al. Downregulation of miR-200a-3p induced by hepatitis B virus X(HBx)protein promotes cell proliferation and invasion in HBV-infection-associated hepatocarcinoma[J]. Pathol Res Pract,2017,213(12):1464-1469.
    [7] HUANG CF,HUANG CY,YEH ML,et al. Genetics variants and serum levels of MHC class I chain-related a in predicting hepatocellular carcinoma development in chronic hepatitis C patients post antiviral treatment[J]. Ebiomedicine,2017,15:81-89.
    [8] HUANG CF,WANG SC,YEH ML,et al. Association of serial serum major histocompatibility complex class I chain-related A measurements with hepatocellular carcinoma in chronic hepatitis C patients after viral eradication[J]. J Gastroenterol Hepatol,2019,34(1):249-255.
    [9] SUN D,WANG X,ZHANG H,et al. MMP9 mediates MICA shedding in human osteosarcomas[J]. Cell Biol Int,2011,35(6):569-574.
    [10] HEIMBACH JK,KULIK LM,FINN RS,et al. AASLD guidelines for the treatment of hepatocellular carcinoma[J]. Hepatology,2018,67(1):358-380.
    [11] Cancer Genome Atlas Research Network. Comprehensive and integrative genomic characterization of hepatocellular carcinoma[J]. Cell,2017,169(7):1327-1341. e23.
    [12] QIN G,DANG M,GAO H,et al. Deciphering the proteinprotein interaction network regulating hepatocellular carcinoma metastasis[J]. Biochim Biophys Acta Proteins Proteom,2017,1865(9):1114-1122.
    [13] CHEUNG KS,SETO WK,WONG DK,et al. Relationship between HBsAg,HBcrAg and hepatocellular carcinoma in patients with undetectable HBV DNA under nucleos(t)ide therapy[J]. J Viral Hepat,2017,24(8):654-661.
    [14] WANG X,OISHI N,SHIMAKAMI T,et al. Hepatitis B virus X protein induces hepatic stem cell-like features in hepatocellular carcinoma by activating KDM5B[J]. World J Gastroenterol,2017,23(18):3252-3261.
    [15] YANG Q,LI XP,ZHONG YB,et al. Interferon-αinhibits cell migration and invasion and induces the expression of antiviral proteins in Huh-7 cells transfected with hepatitis B virus X gene-expressing lentivirus[J]. Exp Ther Med,2017,14(6):5924-5930.
    [16] BRABLETZ T,KALLURI R,NIETO MA,et al. EMT in cancer[J]. Nat Rev Cancer,2018,18(2):128-134.
    [17] SABA NF,WANG Y,FU H,et al. Association of cytoplasmic CXCR4 with loss of epithelial marker and activation of ERK1/2and AKT signaling pathways in non-small-cell lung cancer[J]. Clin Lung Cancer,2017,18(3):e203-e210.
    [18] HAI H,TAMORI A,THUY L,et al. Polymorphisms in MICA,but not in DEPDC5,HCP5 or PNPLA3,are associated with chronic hepatitis C-related hepatocellular carcinoma[J]. Sci Rep,2017,7(1):11912.
    [19] ARAI J,GOTO K,STEPHANOU A,et al. Predominance of regorafenib over sorafenib:Restoration of membrane-bound MICA in hepatocellular carcinoma cells[J]. J Gastroenterol Hepatol,2018,33(5):1075-1081.
    [20] CHEN J,XU H,ZHU XX. Abnormal expression levels of sMICA and NKG2D are correlated with poor prognosis in pancreatic cancer[J]. Ther Clin Risk Manag,2016,12:11-18.
  • Relative Articles

    [1]Tong LIU, Shanzheng LI, Cheng ZHOU, Sutong LIU, Lihui ZHANG, Wenxia ZHAO. Study on the protective efect and mechanism of paeoniflorin on palmitic acid-induced HepG2 cells[J]. Journal of Clinical Hepatology, 2025, 41(3): 499-505. doi: 10.12449/JCH250316
    [2]Lai Xin, Zhou TaiCheng, Wei Jia. Association between HBV A1762T/G1764A double nucleotide substitution and liver diseases[J]. Journal of Clinical Hepatology, 2018, 34(9): 1990-1994. doi: 10.3969/j.issn.1001-5256.2018.09.034
    [3]Pang Ting, Xing HuiChun. Correlation between drug-resistance gene mutations and hepatocellular carcinoma in patients with hepatitis B virus infection[J]. Journal of Clinical Hepatology, 2017, 33(1): 82-86. doi: 10.3969/j.issn.1001-5256.2017.01.017
    [4]Du SenRong, Mao XiaoRong, Xiao Ping, Chen Hong. Effects of PARP-1 inhibitors AG-014699 and AZD2281 on proliferation and apoptosis of human hepatoma cell line HepG2[J]. Journal of Clinical Hepatology, 2015, 31(6): 943-946. doi: 10.3969/j.issn.1001-5256.2015.06.027
    [5]Wang LiPing, Han FangZheng, Li ChunYang, Yan Mei, Zhang YanChao, Yan XueBing. Evaluation of adefovir primary resistance and adefovir repulsive mutations of the HBV DNA P gene in chronic hepatitis B patients[J]. Journal of Clinical Hepatology, 2012, 28(8): 606-608.
    [6]Jin WenWen, Xin YongNing, Dong QuanJiang, Zhao ShouFeng, Yu XinJuan, Jiang Man, Xu Jing, Xuan ShiYing. The preliminary study on the relationship between hepatitis B virus BCP gene mutations and HBV-related primary hepatocellular carcinoma in Qingdao[J]. Journal of Clinical Hepatology, 2012, 28(2): 130-134.
    [7]Yang YanLin, Xiao Ping, Gao Peng, Wang LiMing, Wei XiSheng, He Qiang, Zhou Ping. Distribution of HBV genotypes and YMDD mutations in E antigen-positive patients[J]. Journal of Clinical Hepatology, 2012, 28(6): 428-430.
    [8]Hong YuBo, Zhang Ting, Sun LiGuang. Association between IL-2 +114 (G/T) polymorphism and susceptibility to hepatitis B, cirrhosis, and hepatocellular carcinoma in Jilin province[J]. Journal of Clinical Hepatology, 2012, 28(12): 930-933.
    [9]Deng Le, Wen ZhiLi. Review of recent research progress in correlating HBV genotypes with efficacy of antiviral treatments[J]. Journal of Clinical Hepatology, 2012, 28(6): 469-473.
    [10]Zhang JianJun, Wu LiPing, Du RuiQing, Wang JianBin, Yao ZhanCheng, Yu GuiQin. Factors correlating with the liver inflammation grade ≥G2 in chronic HBV-infected patients with normal liver function[J]. Journal of Clinical Hepatology, 2012, 28(1): 35-38.
    [11]Liu JinPing, Liu YouDe, Guo YanMei, Yang ShaoPing, Lin Qing, Lin Ling. Clinical characteristics and distribution of hepatitis B virus genotypes in CHB patients in Yantai area[J]. Journal of Clinical Hepatology, 2011, 27(12): 1288-1290.
    [12]Zou XiaoJing, Jiang XueQiang, Tian DeYing. Role of DC-SIGN on the maturation and activation of dendritic cells in chronic hepatitis B virus infection[J]. Journal of Clinical Hepatology, 2011, 27(2): 145-147+153.
    [13]Xiong QiXiang, Xiong LiJuan. Hepatitis B virus infection and DNA methylation in hepatocellular carcinoma[J]. Journal of Clinical Hepatology, 2011, 27(8): 881-884.
    [14]Pan Lei, Wei Xin, Xie YuMei, Hao ChunQiu, Wang JiuPing, Jia ZhanSheng, Bai XueFan. Anti-viral therapy with pegylated interferon alpha-2a plus ribavirin in patients co-infected with HBV and HCV[J]. Journal of Clinical Hepatology, 2011, 27(1): 84-85+95.
    [15]Yuan JianGuo, Wang XinGuo, Cui Xia, Liu FengHua, Wang FengHua, Song JiKui, Zhao HongKui. The activation of rat hepatic stellate cells stimulated with hepatitis B virus in vitro[J]. Journal of Clinical Hepatology, 2011, 27(2): 163-165.
    [16]Xi HongLi, Liu FangFang, Yuan JiaYing, Yu YanYan. Significance and detection of IL-2、IL-4 and IFN-γ in PBMC from patients with hepatitis B virus infection[J]. Journal of Clinical Hepatology, 2010, 26(6): 627-629.
    [17]Wang GuoQiang, Niu JuXia. The HBx protein mutation among different HBV genotypes[J]. Journal of Clinical Hepatology, 2010, 26(4): 392-394.
    [18]Shi YanMei, Lan YingHua, Dan Lei, Zhou Jin, Li YongGuo. The relationship between of hematopoietic stem cells and hepatitis B virus infection[J]. Journal of Clinical Hepatology, 2010, 26(4): 377-380.
    [19]Xie DeSheng, Zhang QingBo, Chen ChangXing, Huang JianFang, Ma Jin, Huang HaiBin, Du Yao. The relationship between HBV gene mutation in HBV infected family and HLA class Ⅱ alleles[J]. Journal of Clinical Hepatology, 2010, 26(4): 383-386.
    [20]Xu ZhengJu, Yang Hong, Zhang QiHua, Chen XianLi, Li ShuQing, Wang ChongGuo. Study on the relationship between hepatitis B virus genotypes and effect of lamivudine therapy.[J]. Journal of Clinical Hepatology, 2005, 21(3): 157-159.
  • Cited by

    Periodical cited type(1)

    1. 曾裕文,张芳雍,谭国钳,吴帆. NCEH1基因在肝癌组织与细胞系中的表达及生物学作用. 临床肝胆病杂志. 2021(08): 1867-1872 . 本站查看

    Other cited types(1)

  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Article Metrics

    Article views (1298) PDF downloads(225) Cited by(2)
    Proportional views
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return