关键词 :基因，bcl-2;癌，肝细胞;转染;基因表达

    摘 要:目的 为了探讨抗凋亡基因bcl-2在肝细胞癌(简称肝癌)细胞凋亡过程中的调节功能及其作用机制，建立稳定表达Bcl-2蛋白的肝癌细胞株. 方法 提取含有人bcl-2基因的逆转录病毒真核表达载体pDOR-SB质粒，用限制性内切酶EcoRⅠ(酶切后经琼脂糖凝胶电泳鉴定.用脂质体介导的基因转染法分别将pDOR-SB和pDOR质粒导入不表达Bcl-2蛋白的人肝癌细胞系HCC-9204细胞中，通过G-418筛选获得转入目的基因的阳性细胞克隆.免疫细胞化学法检测Bcl-2蛋白的表达情况.经多次用有限稀释法连续克隆化直至获得100%稳定表达Bcl-2蛋白的肝癌细胞株. 结果 琼脂糖凝胶电泳可见1.9kb的bcl-2基因片断.免疫细胞化学检测表明转染了pDOR-SB的HCC-9204细胞大部分有Bcl-2蛋白的表达，而转染了pDOR空载体或未转染的HCC-9204细胞均未见Bcl-2蛋白表达.经过连续3次克隆化后转染pDOR-SB的HCC-9204细胞100%表达Bcl-2蛋白. 结论 成功地获得了稳定表达Bcl-2蛋白的肝癌细胞株，命名为HCC-bcl2.

    Establishment and identification of the hepatocellu┐lar carcinoma cell strain which expresses Bcl┐2pro┐tein stably

    YANG Lian-Jun，WANG Wen-Liang，SI Xiao-Hui

    1 Department of Pathology，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China，

    2 Institute of Stomatology，Ninth People's Hospital，Shanghai Second Medical University

    Keywords:genes，bcl-2;carcinoma，hepatocellular;transfec-tion;gene expression

    Abstract:AIM A hepatocellular carcinoma(HCC)cell strain which can express Bcl-2protein stably was established to elucidate the regulating function and mechanism of anti-apoptosis gene bcl-2in the apoptotic process of HCC.METHODS The retrovirus eukaryotic expression vector pDOR-SB containing human bcl-2gene was extracted and identified by agarose gel electrophoresis after being cut with restricted endonuclease EcoRⅠ.The pDOR-SB and empty vector pDOR were introduced into a human HCC cell line HCC-9204cells ......