【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 ,为进一步研究HBV和AFB1在肝癌发生过程中的分子机制提供了细胞水平的模型更多还原

【Abstract】 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更多还原