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反义核酸抑制胃癌细胞Bcl-2表达
引用本文:肖冰,王佐佑,时永全,赵燕秋,尤涵,樊代明.反义核酸抑制胃癌细胞Bcl-2表达[J].中国肿瘤生物治疗杂志,1998,5(3):194-198.
作者姓名:肖冰  王佐佑  时永全  赵燕秋  尤涵  樊代明
作者单位:第四军医大学西京医院全军消化病研究所 西安710032
摘    要:本文采用分子克隆技术成功地构建了反义核酸表达载体pDOR-Bcl-2 cDNA.以脂质体介导法将重组反义核酸表达载体转染受体细胞SGC7901,并经C418筛选,从2×10~5细胞中筛选出大约150个抗性克隆,转导率大于1‰,随机挑选2个克隆继续筛选与扩增培养,获得了1株稳定的抗性细胞,从而建立了Bcl-2表达阻断的胃癌细胞株,我们命名为7901anBcl-2 cells.通过Southern印迹法、Northern印迹法、Western印迹法检测显示,转导株与非转导株均有Bcl-2 cDNA的表达,但转导株Bcl-2 mRNA及蛋白的表达明显低于非转导株.说明在胃癌细胞中Bcl-2基因处于高表达状态,通过真核表达载体介导的转染,反义Bcl-2可成功地导入胃癌细胞,并有效地阻断Bcl-2表达.

关 键 词:Bcl-2基因/反义核酸  基因转导  基因表达  胃癌细胞系
收稿时间:1998/4/15 0:00:00
修稿时间:1998/5/11 0:00:00

Expression of Bcl-2 in Gastric Cancer Cells Transducted with Antisense Bcl-2
Xiao Bing,Wang Zuoyou,Shi Yongquan,Zhao Yanqiu,You Han and Fan Daiming.Expression of Bcl-2 in Gastric Cancer Cells Transducted with Antisense Bcl-2[J].Chinese Journal of Cancer Biotherapy,1998,5(3):194-198.
Authors:Xiao Bing  Wang Zuoyou  Shi Yongquan  Zhao Yanqiu  You Han and Fan Daiming
Abstract:By molecular cloning technique, the expressing plasmids pDOR-SV40-Bcl-2 cDNA were successfully constructed. The reconstructed plasmids with lipofectamine were transduced into gastric cancer cell line SGC7901 and then the positive clones which contained the reconstructed plasmids were choosed by G418. Circa 150 positive clones were choosed from 2 x 10_5 gene transduced cells, which suggested that the transducing efficiency was more than 1%o; 2 positive clones were expanded and passage, then 1 drug-resistance cell strain (SGC7901 anBcl-2 cells) was obtained. The bloting results suggested, Bcl-2 cDNA were expressed in both gene transduced and not transduced cell strains, but the expressing level of mRNA and protein in gene transduced cell strain was very low than that in gene not transduced cell strain were positive by the means of Southern blot, Northern blot and Western blot. This results showed that normally in gastric cancer cells the expressing level of Bcl-2 gene was very high, and the cDNA fragment of antisense Bcl-2 were successfully transduced into gastric cancer cells, and in gene transduced cell strains the expressing of Bcl-2 gene was effi-cientlv blocked.
Keywords:Bcl-2 gene/antisense RNA  gene transduction  gene expression  gastric cancer cell
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