慢性乙型肝炎、乙型肝炎肝硬化及原发性肝癌患者HBV-X基因突变分析

雷蔓; 何松

doi:10.3969/j.issn.1001-5256.2014.06.013

慢性乙型肝炎、乙型肝炎肝硬化及原发性肝癌患者HBV-X基因突变分析

DOI: 10.3969/j.issn.1001-5256.2014.06.013

雷蔓,
何松

重庆医科大学附属第二医院消化内科

基金项目:

重庆市卫生局医学科研计划项目重点项目(2013-1-019)；

详细信息

中图分类号: R512.62;R735.7
计量
- 文章访问数: 2441
- HTML全文浏览量: 14
- PDF下载量: 540
- 被引次数: 0
出版历程
- 收稿日期: 2013-09-04
- 出版日期: 2014-06-20

Study of HBV- X gene mutation among patients with HBV- related chronic hepatitis, liver cirrhosis, and primary liver cancer

Lei Man,
He Song

Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University

Research funding:

摘要

摘要: 目的研究慢性HBV感染者,如慢性乙型肝炎（CHB）、乙型肝炎肝硬化（LC）、原发性肝癌（PLC）患者血清中HBV-X基因序列的突变与肝癌发生的关系。方法收集2011-2013年间于重庆医科大学附属第二医院就诊的慢性HBV感染者血清共89例,从血清中提取HBV DNA,扩增全长HBV-X基因序列,经测序后与已知HBV-X基因相应序列比较该患者体内HBV-X基因变异位点以及变异形式,并用卡方检验、单因素方差分析处理数据,NCBI的genotype工具测定基因型。结果所有患者均属于B/C基因型,HBeAg阳性患者中B基因型占46.2%,C基因型占53.8%;HBeAg阴性患性中B基因型占81.2%,C基因型占18.8%（P=0.001）。在PLC组中,启动子（BCP）区的突变显著高于CHB、LC（69.2%vs34.4%和61.3%,P<0.05）,且nt1821位点存在明显的T碱基的缺失（88.5%vs 53.1%和71%,P=0.014）。在CHB、LC中,C基因型BCP的双突变率显著高于B基因型（61.5%vs 15.8%,P=0.007;83.3%vs 47.4%,P=0.04...
- 乙型肝炎,慢性 /
- 肝硬化 /
- 癌,肝细胞 /
- 乙型肝炎病毒 /
- 点突变
Abstract: Objective To study the relationship between hepatocarcinogenesis and the mutation in X gene among patients with chronic hepatitis B virus ( HBV) infection, such as chronic hepatitis B ( CHB) , liver cirrhosis ( LC) and primary liver cancer ( PLC) . Methods The serum samples from 89 patients with chronic HBV infection who visited the Second Affiliated Hospital of Chongqing Medical University from2011 to 2013 were collected. PCR was used to amplify the X gene of HBV DNA extracted from the serum samples. After sequencing, the HBV- X genome was compared with those reported in GenBank to find the variable sites and variant forms. Chi- square and one- way ANOVA were used for the statistical analysis afterwards, whereas genotypes were determined by the genotyping tool of the National Center for Biotechnology Information. Results All patients were genotype B or C. Among HBeAg- positive patients, 46. 2% were genotype B, and53. 8% were genotype C; among HBeAg- negative patients, 81. 2% were genotype B, and 18. 8% were genotype C ( P = 0. 001) . PLC patients had a significantly higher risk of mutation in the basic core promoter ( BCP) region than the CHB and LC groups ( 69. 2% vs 34. 4%and 61. 3%, P < 0. 05) ; in addition, an evident T- base deficiency was observed at nt1821 site ( 88. 5% vs 53. 1% and 71%, P =0. 014) . Among CHB and LC patients, those with genotype C had a significantly higher risk of BCP double mutation than those with genotype B ( 61. 5% vs 15. 8%, P = 0. 007; 83. 3% vs 47. 4%, P = 0. 045) . The incidence of BCP double mutation was significantly higher in the low- viral load group ( ≤106copies /ml) than in the high- viral load group ( > 106copies /ml) ( 81. 3% vs 47. 9%, P = 0. 015) .Conclusion The BCP double mutation and T- base deficiency at nt1821 site may play important roles in the development of PLC.
- hepatitis B, chronic /
- liver cirrhosis /
- carcinoma, hepatocellular /
- hepatitis B virus /
- point mutation

HTML全文

参考文献(32)

[1]WU ZD, WU ZH.Surgery[M].6th ed.Beijing:People's Medical Publishing House, 2002:518-524. (in Chinese) 吴在德, 吴肇汉.外科学[M].6版.北京:人民卫生出版社, 2002:518-524.

[2]FARAZI PA, DEPINHO RA.Hepatocellular carcinoma pathogenesis:from genes to environment[J].Nat Rev Cancer, 2006, 6 (9) :674-687.

[3] GANEM D, PRINCE AM.Hepatitis B virus infection-natural history and clinical consequences[J].N Engl J Med, 2004, 350 (11) :1118-1129.

[4]BLOCK TM, MEHTA AS, FIMMEL CJ, et al.Molecular viral oncology of hepatocellular carcinoma[J].Oncogene, 2003, 22 (33) :5093-5107.

[5]MA L, ZHANG W, XU GZ, et al.Expressions of HBx and CEACAM1 in HBV related hepatocellular carcinoma and their significances[J].J Jilin Univ:Med Edit, 2012, 38 (4) :770-774. (in Chinese) 马莉, 张蔚, 许刚柱, 等.乙型肝炎病毒相关性肝癌患者癌组织中HBx和CEACAM1的表达及其意义[J].吉林大学学报:医学版, 2012, 38 (4) :770-774.

[6]CHEMIN I, ZOULIM F.Hepatitis B virus induced hepatocellular carcinoma[J].Cancer Lett, 2009, 286 (1) :52-59.

[7]KOIKE K.Hepatitis B virus X gene is implicated in liver carcinogenesis[J].Cancer Lett, 2009, 286 (1) :60-68.

[8]KIDD-LJUNGGREN K, MIYAKAWA Y, KIDD AH.Genetic variability in hepatitis B viruses[J].J Gen Virol, 2002, 83 (Pt6) :1267-1280.

[9]KIDD-LJUNGGREN K, MYHRE E, BLACKBERG J.Clinical and serological variation between patients infected with different Hepatitis B virus genotypes[J].J Clin Microbiol, 2004, 42 (12) :5837-5841.

[10]SONG BC, CUI XJ, KIM HU, et al.Sequential accumulation of the basal core promoter and the precore mutations in the progression of hepatitis B virus-related chronic liver disease[J].Intervirology, 2006, 49 (5) :266-273.

[11]BAUMERT TF, MARRONE A, VERGALLA J, et al.Naturally occurring mutations define a novel function of the hepatitis B virus core promoter in core protein expression[J].Virology, 1998, 72 (8) :6785-6795.

[12]SCAGLIONI PP, MELEGARI M, WANDS JR.Biologic properties of hepatitis B viral genomes with mutations in the precore promoter and precore open reading frame[J].Virology, 1997, 233 (2) :374-381.

[13]DATTA S, BANERJEE A, CHANDRA PK, et al.Analysis of hepatitis B virus X gene phylogeny, genetic variability and its impact on pathogenesis:implications in Eastern Indian HBV carriers[J].Virology, 2008, 382 (2) :190-198.

[14]TANAKA Y, MIZOKAMI M.Genetic diversity of hepatitis B virus as an important factor associated with differences in clinical outcomes[J].Infect Dis, 2007, 195 (1) :1-4.

[15]SHINKAI N, TANAKA Y, ITO K, et al.Influence of hepatitis B virus X and core promoter mutations on hepatocellular carcinoma among patients infected with subgenotype C2[J].Clin Microbiol, 2007, 45 (10) :3191-3197.

[16] Chinese Society of Hepatology and Chinese Society of Infectious Diseases, Chinese Medical Association.The guideline of prevention and treatment for chronic hepatitis B (2010 version) [J].J Clin Hepatol, 2011, 27 (1) :Ⅰ-ⅩⅥ. (in Chinese) 中华医学会肝病学分会, 中华医学会感染病学分会.慢性乙型肝炎防治指南 (2010年版) [J].临床肝胆病杂志, 2011, 27 (1) :Ⅰ-ⅩⅥ.

[17]Ministry of Health of the People's Republic of Clina.Diagnosis, management, and treatment of hepatocellular carcinoma (V2011) [J].J Clin Hepatol, 2011, 27 (11) :1141-1159. (in Chinese) 中华人民共和国卫生部.原发性肝癌诊疗规范 (2011年版) [J].临床肝胆病杂志, 2011, 27 (11) :1141-1159.

[18]KUANG SY, JACKSON PE, WANG JB, et al.Specific mutations of hepatitis B virus in plasma predict liver cancer development[J].PNAS, 2004, 101 (10) :3575-3580.

[19]LIZZANO RA, YANG B, CLIPPINGER AJ, et al.The C-terminal region of the hepatitis B virus X protein is essential for its stability and function[J].Virus Res, 2011, 151 (1) :231-239.

[20]LIU XH, LIN J, ZHANG SH, et al.COOH-terminal deletion of HBx gene is a frequent event in HBV-associated hepatocellular carcinoma[J].Gastroenterol, 2008, 14 (9) :1346-1352.

[21] MA NF, LAU SH, HU L, et al.COOH-terminal truncated HBV X protein plays key role in hepatocarcinogenesis[J].Clin Cancer Res, 2008, 14 (16) :5061-5068.

[22]WANG Y, CHEN P, WU X, et al.A new enhancer element, ENII, identified in the X gene of hepatitis B virus[J].Virology, 1990, 64 (8) :3977-3981.

[23]WU X, ZHU L, LI ZP, et al.Functional organization of enhancer (ENII) of hepatitis B virus[J].Virology, 1992, 191 (1) :490-491.

[24]LI M, XIE Y, WU X, et al.HNF3 binds and activates the second enhancer, ENII, of hepatitis B virus[J].Virology, 1995, 214 (2) :371-378.

[25]FAN WM, SHI BY, WEI HS, et al.Comparison of hepatitis B X gene mutation between patients with hepatocellular carcinoma and patients with chronic hepatitis B[J].Virus Genes, 2011, 42 (2) :162-170.

[26]CHEN ST, La PORTE P, YEE JK.Mutational analysis of hepatitis B virus enhancer 2[J].Virology, 1993, 196 (2) :652-659.

[27]PANG A, YUEN MF, YUAN HJ, et al.Real-time quantification of hepatitis B virus core-promoter and pre-core mutants during hepatitis e antigen seroconversion[J].J Hepatol, 2004, 40 (6) :1008-1017.

[28]KAO JH, CHEN PJ, LAI MY, et al.Basal core promoter mutations of hepatitis B virus increase the risk of hepatocellular carcinoma in hepatitis B carriers[J].Gastroenterology, 2003, 124 (2) :327-334.

[29]FANG ZL, YANG J, GE X, et al.Core promoter mutations (A (1762) T and G (1764) A) and viral genotype in chronic hepatitis B and hepatocellular carcinoma in Guangxi, China[J].Virology, 2002, 68 (1) :33-34.

[30]GUO X, JIN Y, QIAN G, et al.Sequential accumulation of the mutations in core promoter of hepatitis B virus is associated with the development of hepatocellular carcinoma in Qidong, China[J].J Hepatol, 2008, 49 (5) :718-725.

[31]LI WH, CHEN GY, YU XW, et al.Accumulation of the mutations in basal core promoter of hepatitis B virus subgenotype C1 increase the risk of hepatocellular carcinoma in Southern China[J].Int J Clin Exp Pathol, 2013, 6 (6) :1076-1085.

[32]CHEN YM, YU DJ, ZHANG WY, et al.HBV subgenotype C2 infection, A1762T/G1764A mutations may contribute to hepatocellular carcinoma with cirrhosis in southeast China[J].Iranian J Public Health, 2012, 41 (11) :10-18.

施引文献

资源附件(0)

访问统计