重组TRAIL基因真核表达载体的构建及其诱导喉癌细胞株Hep2凋亡的效应

目的: 研究TRAIL基因结合端粒酶启动子特异性靶向治疗的作用。方法: 利用已有编码凋亡诱导功能区的TRAILcDNA片断的表达载体pRNDE2 -1和IL- 2信号肽基因序列, 扩增IL- 2信号肽基因和TRAIL基因的融合基因, 并克隆入真核表达载体pGL3 181hTERT肿瘤特异性端粒酶启动子的下游, 构建TRAIL基因的重组真核表达载体pGL3 181hTERT/TRAIL。将该重组载体经阳离子脂质体转染入人喉癌细胞株Hep2中, 以台盼蓝拒染法和基因组DNA琼脂糖凝胶电泳, 观察转染的Hep2细胞的凋亡。结果: 构建了肿瘤人可溶性TRAIL基因的重组真核表达载体pGL3 181hTERT/TRAIL, 其表达产物能诱导喉癌细胞Hep2凋亡。结论: 成功地构建了重组真核表达载体pGL3 181hTERT/TRAIL, 为肿瘤的基因靶向治疗提供了可能性。

Apoptosis of laryngeal carcinoma cell line Hep2 induced by tumor-specific expression of TRAIL gene

AIM: To explore targeted gene therapy of tumor by using the combination of TRAIL gene with the telomerase promoter. METHODS: TRAIL gene with an IL-2 signal peptide was constructed by PCR and cloned into vector pGL3-181hTERT downstream of hTERT promoter to form an eukaryotic expressing vector. Hep2 cells were transfected by the recombinant vector and apoptosis of the transfected cells was evaluated by trypan-blue exclusion and the agarosegel electrophoresis of DNA. RESULTS: We successfully constructed a recombinant eukaryotic expression vector for TRAIL gene.The expressed product significantly induced the apoptosis of Hep2 cells. CONCLUSION: The recombinant eukaryotic expression vector pGL3-181hTERT/TRAIL was successfully constructed, which provides the possibility for gene therapy of tumor.