应用滚环扩增研究肝癌组织中的HBV cccDNA  

Amplification of hepatitis B virus covalently closed circular DNA in patients with hepatocellular carcinoma using rolling circle amplification

作　　者：罗璇[1] 龙泉鑫[1] 黄爱龙[1] 胡源[1] 

机构地区：[1]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016

出　　处：《重庆医科大学学报》2014年第12期1723-1726,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:81101310);教育部博士点新教师基金资助项目(编号:20115503120001);重庆市教委转化成果课题资助项目(编号:Kjzh11201)

摘　　要：目的:运用滚环扩增(rolling circle amplification,RCA)分离肝癌患者组织中乙型肝炎病毒(hepatitis B virus,HBV)共价闭合环状DNA(covalently closed circular DNA,ccc DNA),研究pre-S区的缺失。方法:对RCA技术的特异性和灵敏度进行了研究,并应用该技术扩增13对癌组织和癌旁组织中HBV ccc DNA,对pre-S区的缺失进行了研究。结果:RCA能够高效,特异的扩增组织中10拷贝/μl HBV ccc DNA;26例样本中,22例样本的HBV ccc DNA能够被RCA撤增测序,其中有8例样本(8/22,36.4%)在pre-S区域存在着缺失,pre-S2缺失比例高于pre-S1(8/8 vs.2/8,P=0.007)。结论:首次发现肝癌(hepatocellular carcinoma,HCC)组织中HBV ccc DNA存在着pre-S区的缺失,有助于深入理解ccc DNA pre-S区缺失与HCC的关系。Objective ：To analyze the intrahepatic hepatitis B virus （HBV） covalently closed circular DNA（cccDNA） in hepatocellular carcinoma （HCC） patients. Methods ： The specificity and sensitivity of rolling circle amplification （RCA） were investigated. The intra- hepatic HBV cccDNA in 13 pairs of tumor and adjacent non-tumor tissues were amplified by RCA and pre-S deletions were deter- mined by sequencing. Results ：The HBV cccDNA（ 10 copy/μl） could be amplified specifically by RCA. Among 26 samples, the HBV cccDNA in 22 tissues were amplified successfully, and the pre-S deletions were identified in 8 samples （8/22,36.4%）. In addition, the rate of pre-S2 deletion was higher than that of pre-S1 （8/8 vs. 2/8,P=0.007）. Conclusion：The pre-S2 deletion of HBV cccDNA is first identified in hepatocellular carcinoma patients, which would be benefit for understanding the relationship between cccDNA pre-S deletion and HCC.

关 键 词：乙型肝炎病毒 乙型肝炎病毒共价闭合环状 滚环扩增 肝癌 

分 类 号：R512.62[医药卫生—内科学] R331[医药卫生—临床医学]