摘要
目的探索肝癌(hepatocellular carcinoma,HCC)组织HBV cccDNA T3098C突变情况并检测T3098C突变对HBsAg合成和分泌的影响。方法通过滚环扩增(rolling cycle amplification,RCA)和PCR直接测序,检测23例HCC患者癌组织中T3098C突变情况,并对患者的术后生存时间进行分析,将野生型质粒与T3098C突变质粒分别转染Huh7和HepG2肝癌细胞,检测细胞和上清中HBsAg含量。结果在23例癌组织中,发生T3098C突变的有9例,T3098C突变的生存分析结果显示,C3098突变型组的肝癌患者的生存时间明显长于T3098野生型组,Cox比例风险模型多因素分析发现,T3098C突变是HCC患者术后独立的保护预后预测因素;T3098C突变型组细胞内的HBsAg含量低于HBV 1.2×野生型组(F=42.91,P<0.001),差异具有统计学意义;T3098C突变型组上清和细胞内HBsAg总量低于HBV 1.2×野生型组(F=34.25,P<0.001),差异具有统计学意义。结论T3098C突变是HCC患者独立的保护预后预测因素,这可能与其降低HBsAg的合成和促进HBsAg的外泌有关。
Objective:To explore the existence of HBV cccDNA T3098C mutation in liver cancer tissues and test the effects of T3098C on HBsAg synthesis and secretion.Methods:T3098C mutation in the cancer tissues of 23 HCC patients was detected by rolling cycle amplification(RCA)and PCR direct sequencing,and analyzed with the postoperative survival time of the patients.HBV 1.2 plasmid(HBV C genotype)and HBV 1.2-T3098C mutant plasmid were transfected into Huh7 and HepG2 hepatocellular carcinoma cell lines,respectively,and detected HBsAg content in cells and supernatants.Results:T3098C mutation occurred in 9 of 23 cases of carcinoma tissues,survival analysis results showed that the patients with C3098 had longer survival time than those with T3098,and Cox proportional hazards models multi-factor analysis found that T3098C mutation was the independent protection factor of HCC patients.The experiments also showed that the content of HBsAg in supernatant of T3098C mutant group was lower than that in HBV 1.2 group(F=42.91,P<0.001),the content of total HBsAg of T3098C mutant group was lower than that of HBV 1.2 group(F=34.25,P<0.001).Conclusion:T3098C mutation might be protective factor for HCC patients,which might be related to its decreased HBsAg synthesis and increased HBsAg secretion.
作者
陈忠信
郝泽坤
孙一
虞亚菲
岳宗扬
焦凤萍
CHEN Zhong-xin;HAO Ze-kun;SUN Yi;YU Ya-fei;YUE Zong-yang;JIAO Feng-ping(School of Public Health,Shandong First Medical University&Shandong Academy of Medical Sciences,Taian 271016,China)
出处
《泰山医学院学报》
CAS
2020年第1期5-8,共4页
Journal of Taishan Medical College
基金
国家级大学生创新创业训练计划项目(201810439004)
