乙型肝炎病毒X基因在肝细胞肝癌及慢性乙型肝炎肝细胞染色体上整合情况的对照研究

A compare study of HBx gene integration in hepatocyte chromosome in patients with hepatocellular carcinoma and chronic hepatitis B

目的:应用染色体制备和生物素原位杂交技术,研究乙型肝炎病毒X(HBx)基因在肝细胞肝癌(HCC)细胞染色体上的整合,并与慢性乙型肝炎(CHB)进行对比分析,了解HBx基因在肝细胞染色体上的整合与HCC发生的相关性。同时探讨在HCC与CHB患者中乙肝血清标志物差异及HBx整合与乙肝血清标志物之间的相关性。方法:HCC患者19例,CHB患者14例,分别获取手术或肝穿刺标本并制备成染色体载玻片,以生物素标记的HBx基因探针进行原位杂交,同时统计分析其乙肝血清标志物资料。结果:①19例HCC中有13例患者的肝细胞分裂期染色体上可观察到杂交信号,占68.24%,明显高于CHB组,差别有统计学意义("2=5.12,P<0.05),肝癌患者肝细胞染色体上HBx整合位点数为3.80±1.36,而CHB患者肝细胞染色体上整合位点数为1.30±0.36,二者差别具有极显著性(t=3.5,P<0.01)。②乙型肝炎e抗原(HBeAg)在CHB患者中阳性率为80.0%,明显高于HCC组(10.53%),差别有统计学意义("2=6.33,P<0.01)。③HCC及CHB患者HBx整合与否与患者乙肝血清标志物未见明显相关性。结论:①HBx基因在HCC细胞染色体上的整合明显高于CHB;②CHB患者乙肝病毒复制较HCC患者活跃;③HBx基因整合与否与HCC及CHB乙肝血清标志物未见明显关系。

Objective：To assess the correlation between the HBx gene integration in hepatocyte chromosome and HCC generation, investigate the difference of hepatitis B virus serum markers between patients with HCC and CHB, and to investigate the correlation between HBx gene integration and hepatitis B virus serum markers. Methods：Hepatocyte samples obtained from 19 patients with hepatocyte carcinoma or 14 subjects with chronic hepatitis B were prepared. The frequency of HBx gene integration was detected using biotin-marked HBx probe. Then the results were analyzed combining with the clinical data of hepatitis B virus serum markers. Results： ①Among 19 patients with HCC, 13 subjects were found the signals of HBx gene hybridization in mitotic chromosome of tumour,which occupied 68.24% and was more than that in CHB（28.57%）（X^2 = 5.12 ,P 〈 0.05 ）. The HBx integration sites were 3.8 ：t： 1.36 in hepatocyte chromosome of HCC and 1.3 ± 0.36 in CHB,respectively （t = 3.5,P 〈 0.01）. ②The positive rate of HBeAg in serum was 80.0% in CHB,which is much higher than that in HCC（10.53%）. There was a significant difference（x^2 = 6.33,P 〈 0.01 ） between those two groups. ③There were no correlation between the frequency of HBx integration and the clinical hepatitis B virus serum marks either in HCC or in CHB. Conclusion：The frequency of HBx gene integration in HCC is higher than that in CHB. HBV reproducing in patients with CHB is more active than that with HCC.