肝癌及肝硬化患者血清HBV X基因检测及序列分析

目的:了解肝癌及肝硬化患者血清HBV X基因的表达及其核苷酸序列的变异情况。方法:采用酚-氯仿抽提法抽提患者血清DNA,聚合酶链反应扩增血清HBV X基因,琼脂糖电泳观察结果,并对PCR产物行序列分析。结果:HBV X基因在肝癌患者血清的检出率为32％(16/50),在肝硬化患者血清的检出率为46％(23/50)。经琼脂糖电泳及序列分析证实为HBV X基因。16例肝癌及15例肝硬化患者X基因序列分析显示患者的X基因均存在不同程度的点突变(1～13bp),未发现固定的突变部位与序列。结论:肝癌及肝硬化患者血清HBV X基因存在不同程度点突变,但无特异性突变序列。

Expression and Nucleotide Sequence Analysis of Hepatitis B Virus X Gene in Serum of Hepatocellular Carcinoma and Cirrhotic Patients

Objective: io explore the expression and variety of nucleotide sequence of hepatitis B virus X gene in serum of hepatocellular carcinoma and cirrhotic patients. Methods: DNA was extracted from the serum by phenol-chloroform extracted method and HBV X gene was amplified by polymerase chain reaction. The PCR product was separated in agarose gel and analyzed by automatic sequencing. Results: Serum HBV X gene was found in 32% of HCC patients (16/50) and 46% of cirrhotic patients (23/50). Automatic sequencing analysis showed that different degree of point mutation existed in the X gene (1-13bp), whereas the fixed mutational site and sequence was not obtained. Conclusion: Different degree of point mutations for the X gene exist in hepatocellular carcinoma and cirrhotic patients, whereas the specific mutational sequence has not been discovered.