脂质体介导hES基因转染人舌鳞癌Tca8113细胞及其蛋白表达

Transfection of Human Endostatin Gene with Lipofectamin and the Expression of hES Protein in Tca8113 Cell

目的 建立转人内皮抑素 (hES)基因舌鳞癌Tca8113细胞 ,检测体外培养的转基因细胞基因蛋白表达。方法 为了建立携带hES基因的舌鳞癌Tca8113细胞克隆 ,选用阳离子脂质体Lipofectamin为载体 ,将含hES基因的质粒———pBLAST_hES转染Tca8113细胞 ,以BlasticidinS抗生素筛选阳性克隆。免疫组织化学S_P方法检测体外培养的转基因Tca8113细胞克隆中ES的表达。结果 经BlasticidinS抗生素筛选 ,转染hES基因的Tca8113细胞由于具有bsr抗性基因 ,可以在含有 5 0 μg mlBlasticidinS的培养基中生长和传代 ,从而得到携带hES基因的Tca8113细胞克隆。该细胞在体外培养传代 96h后 ,经免疫组织化学检测 ,hES表达的强阳性 (+++)率为 10 0 %。结论 以脂质体介导hES基因转染Tca8113细胞效率高 ,转基因细胞具有高效表达活性目的基因产物的能力。

Objective The purpose of this study was to establish transfergeneic Tca8113 cell and evaluate the expression of human endostatin (hES) gene in the cell colone in vitro.Methods To transfect hES gene into Tca8113 cells, lipofectamin was complexed with plasmid encoding hES gene, and blasticidin S antibiotic was adopted to select Tca8113-hES cell clone. Immunohistochemistry S-P method was adopted to detect the expression of hES in the transfergenic Tca8113 cell in vitro.Results Transfected by hES, the transfergenic Tca8113 cells could grow and proliferate in RPMI-1640 culture medium containing blasticidin S antibiotic. The expression rate of hES reached 100%.Conclusion hES gene can express in hES-transfected Tca8113 cell in vitro.