地高辛素探针原位杂交用于肝病及肝癌的分子原理研究

Molecular Pathogenesis of Hepatocellular Carcinoma and Chronic Liver Diseases: An in Situ Hybridization Study with Digoxigenin-Labelled Probes

建立了敏感性高、特异性强的地高辛素探针原位杂交技术,并用于肝病和肝癌分子病理学的研究。我们发现慢性乙型肝炎与肝轻微病变52例中43例,肝硬化6例中5例,12例肝癌中10例癌周组织,8例癌组织内可检测出HBV DNA。上述结果揭示了慢性HBV感染与肝癌的分子病理联系。结果还发现原位杂交与转移杂交或PCR检测的结果有较好的一致性。地高辛素探针原位杂交技术可广泛用于消化系疾病分子病理研究。

It is important to detect target genes in pothology specimens of gastroenterological diseases. A sensitive and specific nonradioactive in situ hybridization (ISH) method capable of detecting hepatitis B virus DNA(HBV DNA) sequences in hepatocelluar carcinoma (HCC)and chronic liver disease was developed. The sensitivity of this ISH was sufficient to detect a few copies of Intergrated HBV DNA. The specifity of this method has been proven excellent in double blind control study by southern blot hybridization or polymerase chain reaction. Detection of HBV DNA was carried out in liver tissue from 70 patients. HBV DNA was found in 8 tumous and 10 nontumurous area from 12 patients with HCC, in 5 of the 6 with cirrosis, and 43 of the 52 with chronic liver dieases. This results proved that chronic HBV infection is associated etiolosically with HCC in molecular level. It is concluded that digoxigenin labeling and detection method was highly sensitive and applicable to the detection of viral infection and oncogenes by ISH in gastroenterological diseases.