HBV DNA及HBx DNA亚片段在肝癌细胞内的整合及作用

A study on integration of HBV DNA and HBx subfragment in hepatocellular carcinoma

为探讨 HBV DNA及其亚基因片段 HBx DNA在肝细胞肝癌 ( HCC)中整合特点及作用机制 ,制备了地高辛标记的 3.2 kb HBV DNA全基因及 0 .59kb HBV DNA/ Bam H ,Bgl HBx DNA亚基因探针。以点杂交、原位杂交及 Southern杂交检测 HBV DNA的存在情况 ,筛选 HBV DNA纯整合标本 ,以 HBx DNA探针检测 HBx DNA在 HCC染色体的整合。结果显示 HBV DNA在肝细胞内以核型为主 ( 70 % )。纯整合型占 63.6% ,混合型为 36.4 %。纯整合型中 HBx DNA整合率为 90 .5% ,提示 HBVDNA在 HCC中的整合为普遍存在形式 ,其中 HBx DNA整合在

The purpose of this paper is to study the characteristics of integration of HBV DNA and HBx DNA subfragment in hepatocellular carcinoma (HCC) as well as their effect on the pathogenesis of HCC. Digoxigein labelled probes of 3.2kb HBV DNA and 0.59kb HBV DNA/BamH I Bgl Ⅱ HBx DNA subfragment were prepared. With these probes the presence of HBV DNA were detected using dot blot, in situ hybridization and Southern blot hybridization. The integration rate of HBx DNA subfragment in the chromosomes of HCC cells was subsequencly detected in the specimens which had integrated HBV DNA only. The results showed that HBV DNA localized in liver cells mainly as nucleic type (70%). 63.6% of HBV DNA existed as an integrated type, and 36.4% of HBV DNA was observed in HCC as both integrated and free forms. The integration rate of HBx DNA in the specimens with HBV DNA in pure integration was 90.5%. The result indicated that integration of HBV DNA in HCC was a generally existed form in which the integration of HBx DNA plays an important role in the pathogenesis of HCC.