HBVDNA基因型与肝细胞肝癌的预测性相关研究

将非放射性标记物地高辛甙元用随机引物法由ｐＢＲ３２２－２ＨＢＶ重组质粒／ＥｃｏＲⅠ酶切片段（３．２ｋｂ）制备了乙型肝炎病毒ＤＮＡ探针。用斑点杂交、Ｓｏｕｔｈｅｒｎ杂交、组织原位杂交技术，结合乙肝血清学指标、ｓＩＬ－２Ｒ测定等综合分析了慢性乙型肝炎１７８例及肝细胞肝癌５４例的肝细胞、血清、淋巴细胞、唾液内ＨＢＶＤＮＡ的存在类型、复制表达状态、分布定位特点与病变发展趋势的相关性，探讨了肝癌高危人群肝组织可能发生恶性转化的预测信号及发病机理。

THE RELATIONSHIP RESEARCH OF HBV GENE TYPE WITH THE PREDICTION OF HBPATOCARCINOMA

The main purpose of our study is to bring to light the possible relationship between HBV infection and hepatocarcinoma in molecular level.HBV DNA probe labelled with digoxigenin was prepared by random primer method. One hundred and seventy-eight cases of chronic hepatitis B (CHB) and 54 cases of hepatocarcinoma (HCC) were analyzed by the molecular hybridizaton combined with the serological examinations of HBV markers. The specimens include serum, peripheral blood monocytes (PBMCs) and liver tissue. The results showed: 1.Serum HBV DNA was in a lower level of 35.18% (19/54) in HCC group, and 76.90% (137/178) in CHB group. 2.HBV DNA in liver cells was detected in 88.88% of HCC by dot hybridization, and there was no difference from that in the CHB group (89.28%). The positive rate of serological HBV markers was 93.75% in HCC group. 3.Southern hybridization showed that 65.45% (28/41) of HBV DNA existed in an integrated type and 24.39% (10/41) in both integrated and free types in HCC group. In the CHB group 73.09% (19/26) of HBV DNA existed in free type. 4.The positive rate of HBV DNA was 62.25% (10/16) in PBMCs of HCC patients and 57.40% (8/14) in CHB group. 5.In situ hybridization was used for further confirming the existence of HBV DNA in liver cells.The positive rate of HBV DNA was 80% (24/30), 56.5% of HBV DNA localized in nuclei of the liver cells in HCC patients, and 43.8% in both nuclei and cytoplasm. The results support that there may be two types of HBV infection, integrated and replicated ones.