乙型肝炎病毒X基因变异和慢性乙型肝炎患者临床与组织病理学指标的相关性  

Hepatitis B virus X mutations affect clinicopathological or histopathological characteristics in patients with chronic hepatitis B virus infection

作　　者：包芸[1] 朱虹光[2] 张慧萍[2] 凌玉琴[2] 胡锡琪[2] 朱荣[2] 

机构地区：[1]复旦大学附属华山医院外科病理室,上海200040 [2]复旦大学上海医学院病理学系,上海200032

出　　处：《复旦学报（医学版）》2008年第4期533-538,共6页Fudan University Journal of Medical Sciences

摘　　要：目的探讨慢性乙型肝炎患者肝组织内乙型肝炎病毒（HBV）X基因部分区段的变异与HBV相关各临床与组织病理学指标的关系。方法以83例慢性乙型肝炎肝穿刺活检标本作为研究对象,20例HBV相关性肝细胞癌作为对照;HBV X基因变异的研究采用PCR扩增和直接双向测序。免疫组化二步法显示肝组织内四种病毒蛋白的表达;荧光实时定量PCR检测肝组织内HBV DNA含量。结果HBV X基因nt1583～1793区段错义突变主要出现在对应的X蛋白aa87（nt1632、1633）、88（nt1635、1636）、116（nt1719）、118（nt1726、1727）、119（nt1730）、127（nt1752）、130（nt1762）和131（nt1764）位氨基酸。nt1725～1730（aa118/119）突变与肝组织内HBcAg显著低表达（P=0.006）,和HBV DNA低拷贝数（P=0.004）明显相关,尤以ACTGAC（TD）型突变最为明显;相反,nt1762/1764（aa130/131）位点突变则伴肝组织内HBcAg显著高表达（P=0.005）和高水平的HBV DNA含量（P=0.006）。同时,肝癌组中nt1725～1730野生型所占比率明显高于慢性肝炎（P〈0.05）,而nt1762/1764突变型所占比率肝癌组与肝炎组相比显著增高（P〈0.01）。结论肝组织内,在nt1725～1730突变能显著降低HBcAg的表达和HBV DNA水平,nt1762/1764突变则相反。肝组织内HBV的高负载可能与肝细胞癌的发生有关。Objective To study the relationship between the mutations of hepatitis B virus X gene and the HBV-related clinicopathological or histopathological characteristics in patients with chronic hepatitis B. Methods Eighty-three cases of chronic hepatitis B with liver biopsy were enrolled, and 2（1 HBV-related hepatocellular carcinoma cases were taken as a control. Sequence analysis was performed by polymerase chain reaction （PCR） and the direct sequencing method. Two step immunohistochemical staining showed the expression of four HBV antigens in situ. HBV DNA level in liver was determined by fluorescence quantitative real-time PCR. Results Missense mutations from nt1583 to nt1793 of HBV X gene were detected in position aa87 （nt1632,1633）,88 （nt1635,1636）, 116 （nt1719）,118 （nt1726,1727）,119 （nt1730）,127 （nt1752）,130 （nt1762） and 131 （nt1764）. Mutation at ntl725 - 1730 correlated significantly with the decreased expression of HBcAg in situ （P = 0. 006）, as well as lower HBV DNA levels in liver （P = 0. 004）. Especially CTGAC mutation had the strongest decrease of the viral load （P = 0. 007）. On the contrary, nt1762/1764 mutation correlated with the increased HBcAg （P = 0. 005） and higher HBV DNA levels （P = 0. 006）. The mutants with the wild-type of nt1725 - 1730 or nt1762/1764 mutation were more prevalent in patients with hepatocellular carcinoma than in patients with chronic hepatitis B. Conclusions Mutations at nt1725 - 1730 and nt1762/1764 are associated with in situ expression of HBcAg and viral load. Higher HBV DNA levels in liver is possibly associated with hepatocarcinogenesis.

关 键 词：乙型肝炎 病毒 变异 X基因 

分 类 号：R575.1[医药卫生—消化系统]