重组腺病毒介导Fas基因转染瘢痕疙瘩细胞并诱导其凋亡的实验研究

目的　制备含人Fas基因的重组腺病毒 ,感染瘢痕疙瘩成纤维细胞后 ,使其稳定高效地表达以替换原有无功能的Fas蛋白 ,并恢复正常的重建后Fas信号传导通道。方法　Fas基因自PcDNA3 1 Fas质粒中切取 ,应用PDC315载体系统构建携带Fas基因的重组腺病毒 ,感染瘢痕疙瘩成纤维细胞 ,检测Fas蛋白的表达及功能。结果　成功地构建了携带Fas基因的重组腺病毒 ,感染后瘢痕疙瘩成纤维细胞能稳定表达的有功能的Fas蛋白。结论　利用重组腺病毒转染可以重建被阻断的瘢痕疙瘩成纤维细胞Fas死亡信号传导通道 ,并再次估证了Fas基因突变与瘢痕疙瘩之间的联系 ,为瘢痕疙瘩基因治疗展示了一条新途径

Experimental study on apoptosis of keloid-derived fibroblast transducted with Fas gene by recombinant adenovirus

Objective To generate recombinant adenovirus with human Fas gene and transfect the Fas gene into keloid-|derived fibroblasts to replace the dysfunctional Fas gene for reconstruction of the blocked Fas signal. Methods Fas gene was cut down from the PcDNA3 1 Fas plasmid. The recombinant adenovirus vector was constructed using PDC 3l5 system. Then the expression and function of the transfected Fas gene was detected. Results We successfully constructed the recombinant adenovirus vector with Fas gene and detected its expression and function in the infected keloid|derived fibroblasts. Conclusion We have successfully generated recombinant adenovirus (Ad|PDC315|Fas), which can transfect the Fas gene into keloid|derived fibroblasts to replace the dysfunctional Fas gene and reconstruct the blocked Fas signal.The correlation between the keloid and Fas gene was confirmed.The result may set up a sound foundation for gene therapy of keloid.