乙型肝炎病毒和黄曲霉素协同作用诱发人肝细胞癌细胞株的建立  

Establishment of Human Hepatocellular Carcinoma Cell Line Induced by Synergetic Effect of Hepatitis B Virus and Aflatoxin

作　　者：郭秀婵[1] 蓝祥英[1] 周玲[1] 滕智平[2] 张永利[1] 陈炯玉[3] 沈忠英[3] 曾毅[1] 

机构地区：[1]中国预防医学科学院病毒学研究所,北京100052 [2]北京大学医学部人民医院血液病研究所分子生物学实验室,北京100044 [3]汕头大学医学院肿瘤病理研究室,广东汕头515031

出　　处：《病毒学报》2001年第3期205-209,共5页Chinese Journal of Virology

基　　金：国家自然科学基金重点项目 (3 983 0 3 80 )

摘　　要：为了研究人乙型肝炎病毒 (HBV)和黄曲霉素 (AFB1)在肝癌发生过程中的作用 ,我们用HBV感染的人胚胎肝细胞移植至裸鼠背部皮下 ,以后每周皮下注射AFB1,在裸鼠体内成功地诱发了人肝细胞癌。选 3只裸鼠所形成的肿瘤组织 ,分别再接种裸鼠传代。在裸鼠体内传至 5代后 ,将瘤组织在体外培养、传代 ,建立了 3个肝癌细胞株 ,分别为CBH - 1a、CBH - 1b和CBH - 2。对 3个细胞株进行生物学特性分析发现 ,细胞生长迅速 ,接触抑制消失 ,细胞增殖核计数Ki6 7阳性细胞占 38 2 %。用EMA单抗检测证实为人来源细胞。核酸原位杂交显示 ,细胞中HBV-X和HBV-S基因阳性 ,PCR可扩增出X 基因 ,证明HBV基因已到瘤细胞中。 3个细胞株细胞再接种裸鼠皮下 ,可再生成肿瘤。此实验证明了HBV协同AFB1在人肝细胞癌发生过程中的病因作用。同时 。In order to study the synergetic effect of HBV and aflatoxin in hepatocarcinogenesis,the human embryonic liver cells infected with HBV were transplanted into nude mice by subcutaneous injection Aflatoxin(AFB 1)was injected subcutaneously once a week For the first time,we have successfully induced human hepatocellular carcinomas(HCCs)with HBV and AFB 1 in nude mice Three tumor tissues were separately transplanted into nude mice by subcutaneous injection again After five passages in nude mice the tumor tissues were cultured in vitro and passaged Three human hepatocellular carcinoma cell lines were successfully established and were named CBH-1a,CBH-1b and CBH-2 Analyzing the biological characteristics of these three cell lines,we found that the cells grew rapidly,cell contact inhibition vanished, Ki67 positive cells were 38 2% by counting proliferative cell nuclei The cells were anthropogenic tested by EMA monoclonal antibody The HBV-X and HBV-S genes were demonstrated positive in cell lines by in situ hybridization,PCR amplification revealed X gene It indicated that HBV-X gene had integrated into cellular DNA When cells from three cell lines were transplanted into the nude mice again,the tumor could be induced again Our study demonstrated that there is synergetic effect between HBV and AFB 1 in human hepatocarcinogenesis and it will offer a model on cell level for further research of molecular mechanism of hepatocarcinogenesis

关 键 词：人胚胎肝细胞 乙型肝炎病毒 黄曲霉素 肝癌细胞株 诱导 癌变 协同作用 

分 类 号：R730.231.3[医药卫生—肿瘤] R373.21[医药卫生—临床医学]