宫颈癌Fas阳性表达细胞株的建立

目的　将Fas基因转入宫颈癌细胞 ,建立Fas基因表达株。方法　采用分子克隆技术将Fas基因插入表达载体 pBC的多克隆位点 ,用脂质体介导将Fas基因转入宫颈癌细胞株HeLa中 ,用G4 18筛选克隆细胞 ,扩增并检测Fas基因的表达。结果　成功地构建了表达载体 pBC -FascDNA ,用脂质体介导转入HeLa中 ,可见抗性克隆产生 ,随机挑选 2个克隆扩增培养 ,获得 1株稳定的抗性细胞 ,从而建立了Fas基因表达株 (HeLaFasceells) ,杂交结果表明转导株Fas蛋白表达明显高于非转导株。结论　Fas基因在宫颈癌细胞中处于低表达 ,通过表达载体介导 ,可将Fas基因转入宫颈癌细胞并过表达。

Construction of Fas gene positive cervical carcinoma cell line

Objective To transduct Fas gene into cervical carcinoma cells. Methods The full length cDNA of Fas was inserted into the multiple cloning site of the expressing vector pBC by molecular cloning technique. The reconstructed plasmid with lipofectatmien was transducted into cervical carcinoma cell line HeLa. The positive clones which contained the reconstructed plasmid were chosen by G418. The expression level of Fas gene was determined. Results The expressing plasmid pBC-Fas cDNA was successfully constructed. The postitive clones were generated after the reconstructed plasmid with lipofectamien was transducted into cervical carcinoma cell line HeLa. Two positive clones were randomly chosen and amplified, and 1 drug-resistance cell strain (HeLa Fas cells) was obtained. The blotting results suggested that the expression level of Fas protein was much higher in gene transducted cell strains than those non-transductedones. Conclusion The expression level of Fas gene was very low in cervical carcinoma cells. The cervical carcinoma cells transducted with reconstructed plasmid of pBC-Fas cDNA could express Fas protein efficiently.