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目的将Ｆａｓ基因导入胃癌细胞，建立Ｆａｓ基因表达株，并比较转导前后ｍＲＮＡ与蛋白的表达水平．方法采用分子克隆技术将Ｆａｓ基因插入真核表达载体ｐＢＫＣＭＶ的多克隆克隆位点之间，以脂质体介导法将目的基因导入受体细胞ＳＧＣ７９０１，用Ｇ４１８筛选克隆细胞；以Ｓｏｕｔｈｅｒｎｂｌｏｔ，Ｎｏｒｔｈｅｒｎｂｌｏｔ，Ｗｅｓｔｅｒｎｂｌｏｔ检测Ｆａｓ基因的表达．结果成功地构建了真核表达载体ｐＢＫＦａｓｃＤＮＡ．转导细胞后，从１×１０５细胞中筛选出１００个抗性克隆以上，转导率大于０１％，随机挑选２个克隆扩增培养，获得了１株稳定的抗性细胞，从而有效地建立了Ｆａｓ基因表达株（ＳＧＣ７９０１Ｆａｓｃｅｌｓ）．杂交结果表明，转导株与非转导株均有ｃＤＮＡ的表达，但转导株在ｍＲＮＡ及蛋白水平的表达均明显高于非转导株．结论Ｆａｓ基因在胃癌细胞中处于低表达状态；通过真核表达载体的介导，Ｆａｓ基因导入胃癌细胞后，能有效地表达ＦａｓｍＲＮＡ及其蛋白．
Objective To introduce Fas gene into gastric cancer cells, establish Fas gene expression strain, and compare the mRNA and protein expression levels before and after transduction. Methods The molecular cloning technique was used to insert Fas gene into the cloning site of eukaryotic expression vector pBKCMV. The target gene was introduced into recipient cell SGC7901 by liposome-mediated method, and the cloned cells were selected by G418; Southern blot, Northern blot and Western blot were used to detect the expression of Fas gene. Results The eukaryotic expression vector pBKFascDNA was successfully constructed. After transducing cells, more than 100 resistant clones were screened out from 1×10 5 cells and the transduction rate was greater than 0.1%. Two clones were randomly picked and expanded to obtain a stable resistant cell, which was effective. Fas gene expression strain (SGC7901Fascels) was established. The hybridization results showed that both the transduced and non-transduced strains had cDNA expression, but the transduced strains were significantly higher in mRNA and protein expression than the non-transduced strains. Conclusion The expression of Fas gene is low in gastric cancer cells. Fas gene and its protein can be efficiently expressed in gastric cancer cells mediated by eukaryotic expression vectors.