广西肝癌高发区原发性肝细胞癌 HBV X 基因的整合及影响因素分析

目的 了解广西肝癌高发区乙型肝炎病毒（hepatitis B virus，HBV）X区基因在肝细胞癌（hepatocellular carcinoma，HCC）染色体中的整合及影响因素。方法 以30例与HBV相关的原发性肝细胞癌患者为研究对象。提取HCC组织及癌旁组织标本DNA作为模板，以HBV X基因上游序列和人类基因组Alu重复序列为引物，应用重复序列-多聚酶链反应（Alu-PCR）扩增整合的HBV X片段及两侧的人类基因组DNA片段。扩增产物进行测序，计算目的片段整合率并分析相关的影响因素。结果 18例HCC组织检测到HBV X基因的整合片段，整合率为60.00%（18/30）；26例癌旁组织检测到HBV X基因的整合片段，整合率为86.67%（26/30）。癌旁组织HBV病毒整合率高于HCC组织，差异有统计学意义（χ²=5.445，P=0.020）。不同性别、年龄、HBeAg、HBV DNA、ALT、AST的HCC癌组织及其癌旁组织HBV X基因整合率比较差异均无统计学意义（P＞0.05）。结论 广西肝癌高发区癌旁组织比HCC组织HBV整合率高，说明HBV整合发生在感染早期。HBV X基因整合与HCC患者性别、年龄、HBeAg、HBV DNA、ALT、AST无明显关系。

Analysis of hepatitis B virus X gene integration and potential risk factors of integration in patients with hepatocellular carcinoma in the high-incidence area of Guangxi

Objective To analyze integration of hepatitis B virus（HBV） X gene into the genome of primary hepatocellular carcinoma （HCC） tumors and surrounding tissue，and to explore potential risk factors of such integration in patients from the HCC high-incidence area of Guangxi. Methods DNA was extracted from HCC tissue and adjacent non-tumor liver tissue of 30 patients with HBV-related HCC. Primers were designed to amplify the HBV X sequence and human Alu repeats by Alu-PCR，and PCR products were Sanger- sequenced. Results Among the 30 patients，18（60.00%） showed integration of the HBV X gene into HCC tumors，compared to 26 （86.67%） who showed X gene integration into adjacent liver tissue （χ²=5.445，P=0.020）. Incidence of X gene integration did not vary significantly with sex，age，HbeAg status，HBV DNA load，ALT or AST in HCC tissue or adjacent non-tumorous liver tissue（P>0.05）.Conclusions Incidence of HBV integration is higher in non-tumorous tissues than in tumorous tissues in patients with HBV-related HCC in Guangxi.HBV integration appears to occur in early stages of HCC development. HBV X gene integration does not appear to be influenced by sex，age，HBeAg status，HBV DNA load，ALT or AST.