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
作者：潘朝斌,黄洪章,王建广,侯劲松
Author：PAN Chao-bin,HUANGHong-zhang,WANGJian-guang,HOUJin-song
收稿日期：2004-04-25 年卷（期）页码：2004,22(02):96-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：内皮抑素,转基因,细胞,舌鳞癌,
Key words：endostatin,gene transfer,cell,squamous cell carcinama of tongue,
基金项目：国家自然科学基金资助项目(30271423);广东省医学科学技术研究基金(2003A025)
中文摘要
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Blasticidin S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Blasticidin S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50μg/ml Blasticidin S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96 h后,经免疫组织化学检测,hES表达的强阳性(+++)率为100%。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。
英文摘要
Objective The purpose of this studywas to establishtransfergeneicTca8113 cell and evaluate the expression of hu- man endostatin (hES) gene in the cell colonein vitro.Methods To transfect hES gene into Tca8113 cells, lipofectamin was complexed with plasmid encoding hES gene, and blasticidin S antibiotic was adopted to selectTca8113-hES cell clone. Immuno- histochemistry S-P method was adopted to detect the expression of hES in the transfergenic Tca8113 cellin vitro.Results Trans- fected 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 cellin vitro.
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