HBV亚基因整合及其致癌性研究

HBV SUBGENE INTEGRATION AND ITS CARCINOGENICITY

目的探讨乙型肝炎病毒亚基因在肝细胞癌发生中的作用。方法以ＢａｍＨＩ、ＢｇｌⅡ酶切３．２ｋｂＨＢＶＤＮＡ，回收其ＨＢｓ、ＨＢｘ、ＨＢｃ、ＰｒｅＳ亚基因片段，以地高辛甙元标记成探针。以ＨＢＶ探针Ｓｏｕｔｈｅｒｎ杂交分析其存在状态，以ＨＢＶ亚基因探针Ｎｏｒｔｈｅｒｎ杂交检测亚基因转录。结果ＨＢＶ阳性肝细胞癌中，整合型与混合型标本分别为６３．６％和３６．４％，纯整合型标本中，ＰｒｅＳ、ＨＢｓ、ＨＢｘ、ＨＢｃｍＲＮＡ阳性率分别为１１．１％、４４．４％、５５．６％、１１．１％。结论ＨＢＶＤＮＡ以整合于染色体上作为存在的主要方式，整合的ＨＢＶＤＮＡ有一定程度的转录，指导相应蛋白合成而影响肿瘤的发生。

Objective To study the action of HBV subgenes in hepatocarcinogenesis.Methods HBV DNA 3.2 kb was digested with BamH Ⅰ and Bgl Ⅱ to retrive HBs, HBx, HBc and PreS subgene fragments, which were then labelled with digoxigenin to form probes. The status of HBV existance was tested with HBV probe by Southern blot hycridization and the subgene transcription was examined by Northern blot hybridization separately using HBV subgene probes.Results In HBV positive hepatocellular carcinoma (HCC) specimens, th integrated type and the mixed type of HBV accounted for 63.6% and 36.4% respectively. In the integrated type specimens, the mRNA positive rates of PreS, HBs, HBx and HBc were 11.1%,44.4%,55.6% and 11.1% respectively.Conclusion HBV DNA integrates itself into the chromasome as the main form of its existance in HCC cells. The integrated HBV DNA may be partly transcribed to alter protein synthesis and play certain roles in hepatocarcinogenesis.