L02细胞乙型肝炎病毒X蛋白和SOCS-1基因水平研究*

doi:10.3969/j.issn.1672-5069.2022.06.004

实用肝脏病杂志 ›› 2022, Vol. 25 ›› Issue (6): 772-775.doi: 10.3969/j.issn.1672-5069.2022.06.004

L02细胞乙型肝炎病毒X蛋白和SOCS-1基因水平研究^*

贺潇瑾, 李丹, 周青, 欧阳静, 田玉球, 谭英征

412006 湖南省株洲市中南大学湘雅医学院附属株洲医院感染内科

收稿日期:2022-03-29 出版日期:2022-11-10 发布日期:2022-11-22
通讯作者: 谭英征,E-mail:346370009@qq.com
作者简介:贺潇瑾,女,31岁,硕士研究生,主治医师。主要从事病毒性肝炎与肝癌发生研究
基金资助:
^*湖南省科技创新计划项目(编号:2020SK53901)

Expression of hepatitis B virus X protein and suppressor of cytokine signaling-1 in L02 cells in vitro

He Xiaojin, Li Dan, Zhou Qing, et al.

Department of Infectious Diseases, Zhuzhou Hospital Affiliated to Zhongnan University Xiangya Medical School, Zhuzhou 412006, Hunan Province, China

Received:2022-03-29 Online:2022-11-10 Published:2022-11-22

摘要/Abstract

摘要： 目的探讨乙型肝炎病毒X蛋白(HBx)影响细胞因子信号传导抑制因子-1(SOCS-1)基因水平的可能机制。方法收集22例乙型肝炎相关肝细胞癌(HCC)手术后癌组织和癌旁肝组织,采用RT-PCR法检测组织HBx、DNMT3A、DNMT3B和SOCS-1 mRNA水平。通过脂质体法构建表达HBx的L02细胞和空质粒L02细胞,采用CCK8法检测5-氮杂-2′-脱氧胞苷(5-Aza-c)对L02细胞存活率的影响,采用RT-PCR法和Western blot法分别检测细胞HBx、DNA甲基转移酶(DNMT)3A、DNMT3B和SOCS-1 mRNA水平及其蛋白表达。结果肝癌组织HBx和DNMT3A mRNA水平分别为(65.2±3.5)和(77.2 ± 3.8),显著高于癌旁肝组织【分别为(22.5±4.0)和(42.1± 2.9),P<0. 05】,而SOCS-1 mRNA水平为(33.1±3.0),显著低于癌旁肝组织【(75.6 ±2.6),P<0. 05】;与对照细胞比,表达HBx的L02细胞活力随5-Aza-c作用浓度增高而下降(P<0. 05);过表达HBx的L02细胞DNMT3A mRNA水平及其蛋白表达量显著高于空载质粒组(P<0.05),而SOCS-1 mRNA水平及其蛋白表达量显著低于空载质粒组(P<0. 05);在5-Aza-c干预表达HBx的L02细胞,DNMT3A mRNA水平及其蛋白表达量显著低于对照组(P<0. 05),而SOCS-1 mRNA水平及其蛋白表达量显著高于对照(P<0. 05)。结论 HBx通过上调DNMT3A表达使SOCS-1基因表达下调,表明HBx可能通过调控DNMT3A影响抑癌基因SOCS-1表达从而促进肝癌的发生。

关键词: 肝细胞癌, L02细胞, 乙型肝炎病毒X蛋白, 细胞因子信号传导抑制因子-1, DNA甲基化转移酶, 体外

Abstract: Objective The aim of this study was to investigate the possible mechanism of hepatitis B virus X protein (HBx) affecting cytokine signal transduction inhibitor-1 (SOCS-1) gene in vitro. Methods The expressions of HBx, DNA methyltransferase (DNMT)3A/3B and SOCS-1 in cancerous and paracancerous tissues of 22 patients with HBV-related hepatocellular carcinoma (HCC) were detected by real-time PCR. The HBx expression plasmid (pcDNA-X) or an empty plasmid (pcDNA3. 0) were transfected in L02 cells by liposome infection. The effect of 5-aza-2′-deoxycytidine(5-Aza-C) on the survival rate of L02 cells was detected by CCK8. The HBx, DNMT3A/3B and SOCS-1 mRNA as well their protein expression were assayed by real-time PCR and Western blot. Results The HBx and DNMT3A mRNA level in cancerous tissues were(65.2±3.5)and (77.2 ± 3.8), much higher than [(22.5±4.0)and(42.1± 2.9), respectively, P<0. 05], while the expression of SOCS-1 was (33.1±3.0), significantly lower than [(75.6 ±2.6),P<0. 05] in adjacent liver tissues; the activity of L02 cells expressing HBx decreased with the increase of 5-Aza-C concentration (P<0. 05); the DNMT3A mRNA level and its protein expression in L02 cells with overexpression of HBx were significantly higher than in empty plasmid-transfected cells (P< 0.05); the SOCS-1 mRNA level and its protein expression were significantly lower than in the empty plasmid-transfected cells (P< 0.05); the DNMT3A mRNA level and its protein expression in L02 cells expressing HBx after 5-Aza-C intervention were significantly lower than in the control cells (P < 0.05), while the SOCS-1 mRNA level and its protein expression were significantly higher than in the control (P < 0.05). Conclusions The present study indicates that HBx induces epigenetic down-regulation of SOCS-1 by increasing the expression of DNMT3A,which might be reversed with DNA methyltransferase inhibitor 5-Aza-C.

Key words: Hepatoma, L02 cells, Hepatitis B virus X protein, Suppressor of cytokine signaling-1, DNA methylation transferase, In vitro

贺潇瑾, 李丹, 周青, 欧阳静, 田玉球, 谭英征. L02细胞乙型肝炎病毒X蛋白和SOCS-1基因水平研究^*[J]. 实用肝脏病杂志, 2022, 25(6): 772-775.

He Xiaojin, Li Dan, Zhou Qing, et al.. Expression of hepatitis B virus X protein and suppressor of cytokine signaling-1 in L02 cells in vitro[J]. Journal of Practical Hepatology, 2022, 25(6): 772-775.

参考文献

[1] Kulik L, El-Serag HB. Epidemiology and management of hepatocellular carcinoma. Gastroenterology, 2019,156(2):477-491,e1.
[2] 严景全,王春玲,刘娟,等.血清乙型肝炎病毒前基因组RNA水平对核苷(酸)类似物初治的慢性乙型肝炎患者疗效预测价值.实用肝脏病杂志,2022,25(1):22-25.
[3] 孙海荣,冯伟广,周冰清,等.不同血清ALT水平的乙型肝炎病毒感染者肝纤维化指标变化.实用肝脏病杂志,2021,24(4):488-491.
[4] Nebbioso A, Tambaro FP, Dell'Aversana C, et al. Cancer epigenetics: moving forward. PLoS Genet, 2018,14(6):e1007362.
[5] Zhao RC, Zhou J, He JY , et al. Aberrant promoter methylation of socs-1 gene may contribute to the pathogenesis of hepatocellular carcinoma: a meta-analysis. J BUON, 2016,21(1):142-151.
[6] Köhler F, Rodríguez-Paredes M. DNA methylation in epidermal differentiation, aging, and cancer. J Invest Dermatol, 2020, 140(1):38-47.
[7] Lyko F.The dna methyltransferase family: a versatile toolkit for epigenetic regulation.Nat Rev Genet, 2017,19(2):81-92.
[8] Gujar H, Weisenberger DJ, Liang G. The roles of human DNA methyltransferases and their isoforms in shaping the epigenome. Genes (Basel), 2019, 10(2):172.
[9] Ribatti D, Tamma R. Epigenetic control of tumor angiogenesis. Microcirculation, 2020,27(3):26-32.
[10] iang S, Guo Y. Epigenetic clock: DNA methylation in aging. Stem Cells Int, 2020,10(4):47-68.
[11] Loaeza-Loaeza J, Beltran AS, Hernández-Sotelo D. DNMTs and impact of CpG content, transcription factors, consensus motifs, lncRNAs, and histone marks on DNA methylation. Genes (Basel), 2020,11(11):13-23.
[12] 李引钰,周易.乙型肝炎病毒X蛋白相关lncRNA NEAT1促肝癌增殖和转移的研究.中国病原生物学杂志,2021,16(10):1122-1128.
[13] 黎蓓,马艳,李盈,等.肝癌组织中SOCS1、GSTP1及DKK3基因甲基化状态及与临床病理的关系.解放军医药杂志,2021,33(10):37-40,53.
[14] Xie Q, Chen L, Shan X, et al. Epigenetic silencing of sfrp1 and sfrp5 by hepatitis B virus X protein enhances hepatoma cell tumorigenicity through WNT signaling pathway. Int J Cancer, 2014, 135(3):635-646.
[15] Zhao J, Wu G, Bu F, et al. Epigenetic silence of ankyrin-repeat-containing, sh3-domain-containing, and proline-rich-region- containing protein 1 (aspp1) and aspp2 genes promotes tumor growth in hepatitis b virus-positive hepatocellular carcinoma. Hepatology, 2010, 51(1):142-153.
[16] Wang X, Yan J. Effect of Hbx-mediated RXR alpha repression on dnmt3a in hepatocellular carcinoma and underlying mechanism. J Third Milit Med Uni, 2017,22(1):91-99.
[17] Wei X, Xiang T, Ren G, et al. Mir-101 is down-regulated by the hepatitis B virus X protein and induces aberrant dna methylation by targeting DNA methyltransferase 3a. Cell Sig, 2013,25(2):439-446.
[18] Zhao RC, Zhou J, He JY, et al. Aberrant promoter methylation of socs-1 gene may contribute to the pathogenesis of hepatocellular carcinoma: a meta-analysis. J BUON, 2016,21(1):142-151.
[19] Pazienza V , Panebianco C , Andriulli A. Hepatitis viruses exploitation of host DNA methyltransferases functions. Clin Exp Med, 2016, 16(3):265-272.
[20] Fernández-Barrena MG, Arechederra M, Colyn L, et al. Epigenetics in hepatocellular carcinoma development and therapy: the tip of the iceberg. J Hep Rep, 2020, 2(6):100167.

L02细胞乙型肝炎病毒X蛋白和SOCS-1基因水平研究^*

Expression of hepatitis B virus X protein and suppressor of cytokine signaling-1 in L02 cells in vitro

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