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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 study the integration of HBx gene on the chromosome of hepatocellular carcinoma (HCC) cells by chromosomal preparation and biotin in situ hybridization and to compare with chronic hepatitis B (CHB) Correlation of HBx Gene Integration on Hepatocyte Chromosomes with HCC Occurrence. At the same time to explore the difference between hepatitis B serum markers and HBx integration in patients with HCC and CHB serum hepatitis B serum markers. Methods: Twenty-nine patients with HCC and 14 patients with CHB were included in this study. Surgical or hepatic puncture specimens were obtained and prepared into chromosomal slides. HBs gene probes labeled with biotin were used for in situ hybridization. Meanwhile, the serum HBV markers . Results: (1) The signal of hybridization was observed on the chromosomes of hepatocytes in 13 of 19 HCC patients, accounting for 68.24%, which was significantly higher than that of CHB group (P <0.05) The number of HBx integration sites on the chromosome of hepatocellular carcinoma was 3.80 ± 1.36, while the number of integration sites on the chromosome of hepatocellular carcinoma was 1.30 ± 0.36 in CHB patients (t = 3.5, P <0.01) The positive rate of hepatitis B e antigen (HBeAg) in CHB patients was 80.0%, which was significantly higher than that in HCC patients (10.53%), the difference was statistically significant ( "2 = 6.33, P <0.01). ③HCC and CHB HBx patients with or without integration of hepatitis B serum markers and no significant correlation. Conclusion: ①HBx gene integration in the chromosome of HCC cells was significantly higher than CHB; ② CHB patients with HBV replication than HCC patients active; ③ HBx gene integration or not and HCC and CHB hepatitis B serum markers no significant relationship.