携带Fas基因重组腺病毒治疗瘢痕疙瘩的体外研究

EXPERIMENTAL GENE THERAPY OF KELOID IN VITRO USING RECOMBINANT ADENOVIRUS CODING FOR FAS GENE

目的 应用成功制备的携带人 Fas基因的两种重组腺病毒 ,感染瘢痕疙瘩 (keroid,K)成纤维细胞 (fi-broblasts,FB) ,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 腺病毒感染 KFB后 ,检测及比较两种腺病毒转染前后 ,KFB内 Fas蛋白的表达变化、蛋白功能以及细胞增殖 -凋亡状况。 结果 腺病毒感染后的 KFB均能稳定提高 Fas蛋白的表达 ,并且在 Fas单克隆抗体的诱导下可以发生明显的细胞凋亡。 结论 1构建的两种重组腺病毒 Ad- Fas在体外实验中能有效地提高 KFB蛋白的表达 ,并可以重建原来阻断的死亡信号传导通道 ;2细菌内重组腺病毒体外治疗效果更佳 ;3再次估证了 Fas基因突变与 K之间的联系 ,为 K基因治疗展示了一条新途径。

Objective To replace dysfunctional Fas gene and reconstruct the blocked Fas signal by using two kinds of prepared recombinant Adenovirus which have human Fas gene . Methods After the keloids derived from fibroblasts were infected by the Adenovicus, the expressions of Fas protein before the exposure and after the exposure was compared. Then the function of the newly produced Fas protein was detected. Results The highly improve expression of Fas protein in the infected keloid derived fibroblasts was detected. Obvious apoptosis was also detected in the infected keloid derived from fibroblasts under the condition of exposing to FasMcab. Conclusion ①The recombinant Adenovirus with Fas gene can transfect the Fas gene into keloid derived fibroblasts and highly improved the expression of Fas protein. The newly expressed Fas gene can reconstruct the blocked Fas signal. ②Ad Fas(B) has better therapeutic effect in vitro gene therapy. ③ The correlation between keloid and Fas gene was further proved and it may pave the way for further gene therapy in keloid .