| L02细胞乙型肝炎病毒X蛋白和SOCS-1基因水平研究* |
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| 引用本文： | 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. |
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| 作者姓名： | 贺潇瑾 李丹 周青 欧阳静 田玉球 谭英征 |
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| 作者单位： | 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 |
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| 基金项目： | *湖南省科技创新计划项目(编号:2020SK53901) |
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| 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。
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| 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 |
| 收稿时间： | 2022-03-29 |
Expression of hepatitis B virus X protein and suppressor of cytokine signaling-1 in L02 cells in vitro |
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| 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 Clinical Hepatology,2022,25(6):772-775. |
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| Authors: | He Xiaojin Li Dan Zhou Qing |
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| Affiliation: | Department of Infectious Diseases, Zhuzhou Hospital Affiliated to Zhongnan University Xiangya Medical School, Zhuzhou 412006, Hunan Province, China |
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| 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. |
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| Keywords: | Hepatoma L02 cells Hepatitis B virus X protein Suppressor of cytokine signaling-1 DNA methylation transferase In vitro |
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