肝细胞癌患者石蜡包埋肝组织HBV ccc DNA的检测  

Detection of HBV ccc DNA in FFPE liver tissues of patients with hepatocellular carcinoma

作　　者：胡双烨[1] 徐晨[2] 赵雨来[2] 李智彬[2] 徐东平[2] 刘永明[1] 杨媛[2] 赵景民[2] 成军[3] 苏何玲[1] 侯俊[2] 钟彦伟[2] 

机构地区：[1]桂林医学院,桂林市541004 [2]解放军第302医院全军传染病研究所 [3]首都医科大学附属北京地坛医院

出　　处：《中华实验和临床感染病杂志（电子版）》2013年第2期5-8,共4页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：首都医学发展科研基金(No.2009-3057)

摘　　要：目的检测肝细胞癌患者肝组织中乙型肝炎病毒共价闭合环状DNA(HBV ccc DNA),探讨其在肝细胞癌发生、发展中的意义。方法选取20例肝细胞癌患者的癌及癌旁组织,经10%福尔马林固定、石蜡包埋、连续切片,采用0.05%多聚赖氨酸进行处理。经不降解质粒的ATP依赖的DNA酶(PSAD酶)消化后,应用原位滚环扩增(insitu RCA)结合原位跨缺口PCR扩增技术检测肝组织中ccc DNA的表达。结果 20例肝癌患者标本中有17例检测到HBV ccc DNA阳性信号,为蓝色或紫蓝色并呈团块状,定位于细胞核。肝癌患者癌及癌旁组织HBV ccc DNA阳性率分别为10%(2/20)和85%(17/20),差异具有显著统计学意义(P<0.01)。结论肝细胞癌患者癌旁组织病毒复制水平高于癌组织。Objective To detect the intrahepatic covalently closed circular DNA （ccc DNA） in patients with HBV-related hepatocellular carcinoma （HCC） and to investigate its significance in progression of HCC. Methods The paired tumor and adjacent non-tumor tissues of 20 cases with HCC were selected, respectively. The samples were fixed by 10% formaldehyde, paraffin imbedded and treated with 0.05% poly- L-Lysine. The liver tissue sections were treated by plasmid-safe ATP-dependent DNase （PSAD） and the expression of intrahepatic ccc DNA was detected by in situ rolling circle amplification （RCA） combined with in situ polymerase chain reaction （PCR） targeting the gap region. Results HBV ccc DNA was expressed in 17 of 20 subjects and located in the nuclear compartment with clustering, staining blue or blue-violet. The positive rate of HBV ccc DNA in HCC and their adjacent tissues were 10% （2/20） and 85% （17/20）, respectively, with significant difference （P 〈 0.01）. Conclusion The level of HBV replication in the tumoradjacent tissues was higher than that of the tumor tissues of patients with HCC.

关 键 词：肝炎病毒 乙型 共价闭合环状DNA 肝细胞癌 

分 类 号：R735.7[医药卫生—肿瘤]