乙型肝炎病毒（HBV）感染是导致原发性肝癌的重要原因。HBV整合是致癌的可能机理之一。本研究拟应用HBV-Alu-PCR方法筛选我国肝癌患者癌组织标本，分离鉴定HBVDNA在宿主细胞染色体上的整合位点，分析位于HBVDNA侧翼的细胞基因序列和类型，并对所获新基进行结构和功能的研究，为进一步探索HBV致癌机理提供新思路。

In order to study the relationship of hepatitis B virus (HBV) integration and hepatocellular carcinogenesis, a rapid method based on Alu-PCR technic was established to identify the viral-cellular junction. By using HBV-Alu-PCR method, we successfully characterized 68 HBV integration sites from 40 hepatocellular carcinoma(HCC). Eighty-five percent of HBsAg-positive Chinese HCC samples showed at least one copy of integrated HBV DNA in host genomic. Viral DNA most frequently inserted into host chromosome 2, 3, 19 and 22, whereas chromosome 10、15、16、18、20、21、X and Y did not contain any HBV insertion. HBV preferred to integrate into the intron or upstream regulatory region of tumor-related genes. There are 4 genes, i.e. myeloid/lymphoid or mixed-lineage leukemia 4 （MLL4）, G protein alpha transducing activity polypeptide 1 （GNAT1）, fibronectin 1（FN1）and mitogen-activated protein kinase 1(MAPK1) , found to be repeatedly targeted by HBV. Different from the previous report that HBV integrates into host genome at DR1 and DR2 regions, our results clearly showed that viral genome could be interrupted at any position of HBV X gene. In 96% cases, X gene was deleted at 3'-end, which supported our hypothesis that N-terminal truncated X protein may play a more important role in cell transformation than full-length HBx. The resul

在国内首次建立了高效、准确分离乙肝病毒（HBV）整合位点的HBV-Alu-PCR方法，并以该方法对我国40例肝癌组织标本进行了HBV整合位点分析。 结果显示我国乙肝表面抗原阳性的肝癌组织中，85%有HBV DNA整合；病毒整合多发生于2、3、19和22号染色体，10、15、16、18、20、21、X和Y号染色体未见病毒整合；HBV偏好插入肿瘤相关基因的内含子和上游调控区；并发现四个基因（MLL4,GNAT1,FN1,MAPK1）被HBV重复锚定。上述结果表明HBV在肝癌细胞基因组中的整合具有一定的规律。此外，从病毒方面分析，整合可发生于X基因的任何长度，并不集中于通常认为的DR1和DR2区，但在96%的标本中， X基因均以截短形式插入宿主细胞DNA ，提示了截短型X蛋白的反式致癌作用。本研究是迄今为止国内外所进行的最大规模的有关HBV整合位点的筛选，其结果不仅导致再次发现了被HBV重复整合的基因，而且使我们以整合为线索，克隆了6个可能与肝癌相关的新基因（C13orf10, C22orf15, KIAA0515, KIAA0888, LOC441054, MGC39518）。