肝细胞癌患者石蜡包埋肝组织HBVcccDNA的原位检测-病理学与病理生理学专业论文.docx

PAGE PAGE 2 肝炎患者肝组织3例）作为阴性对照组。结果：20例肝细胞癌患者肝组织标 本中有17例（85%）检测到HBVcccDNA阳性信号，为紫红色或紫蓝色呈团 块状,定位于细胞核。肝癌患者癌组织及癌旁组织HBVcccDNA阳性率分别为 10%(2/20)和85% (17/20) , 两者差异有显著性(P0.01)。阴性对照组均未见阳 性信号表达。结论：PSAD酶消化加滚环扩增结合原位PCR方法能够定位检 测肝癌组织和癌旁组织中HBV cccDNA的表达，该方法具有一定的特异性和 敏感性；肝细胞癌患者癌旁组织病毒复制水平高于癌组织。 关键词：乙型肝炎病毒；共价闭合环状 DNA;肝细胞癌；原位 PCR；滚 环扩增 The in situ detection of HBVcccDNA in FFPE liver tissues of patients with hepatocellular carcinoma ABSTRACT Objective: Primary hepatolcellular carcinoma（HCC）is one of the common highly malignant tumors, with second highest incidence for all cancers in China. Epidemiology has confirmed that infection of HBV is the independent risk of HCC. Covalently closed circular DNA (cccDNA) is an important replicative intermediate in the life period of HBV, which serves as the HYPERLINK /dict_result.aspx?searchword=%e5%8e%9f%e5%a7%8bamp;tjType=sentenceamp;styleamp;t=original original template for viral and pregenomic messenger RNA (pgRNA),has been generally considered as the basic reason of persistent viral infection and liver progressive pathological changes. Studies in vitro and animal researchs have shown that there is a closely relationship between the HBVcccDNA and hepatocellular carcinoma. With the PCR development, quantitative detection techniques of intrahepatic cccDNA have been matured. But there are less reports about in situ detection of cccDNA from liver tissues of patients with HCC domestic and overseas. Therefore, we developed a novel in situ method to detect the intrahepatic cccDNA in tumor and adjacent non-tumor tissues in HCC patients and to investigate the significance in HCC progression and provide theory for prevention and treatment of primary liver cell carcinoma.Methods : The paired tumor and adjacent non-tumor tissues tumor tissues were obtained from 20 HCC patients hospitalized in Beijing 302 Hospital during July 2011-July 2012. The samples were fixed by 10% formaldehyde, paraffin imbedded and serial sectioned at 5μm. The liver tis