稳定表达Bcl┐2蛋白的肝细胞癌细胞株的建立与鉴定.doc

稳定表达Bcl┐2蛋白的肝细胞癌细胞株的建立与鉴定
作者:杨连君,王文亮,司晓辉
【关键词】基因
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(-SB和pDOR质粒导入不表达Bcl--9204细胞中,通过G--%稳定表达Bcl-2蛋白的肝癌细胞株. ---9204细胞大部分有Bcl-2蛋白的表达,-9204细胞均未见Bcl---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,-bcl2.

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 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,which do not express Bcl-2protein,by lipo-some-mediated gene transfection cell clones into which pDOR-SB or pDOR had been introduced were obtained with G- expression of Bcl-2protein was detected using immunocytochemical pDOR-SB-tranfected cells were