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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。
Objective: To investigate the role of TRAIL gene in combination with telomerase promoter-specific targeted therapy. Methods: The fusion gene of IL-2 signal peptide and TRAIL gene was amplified by using the sequence of the expression vector pRNDE2 -1 and IL-2 of the TRAIL cDNA fragment encoding the apoptosis-inducing functional domain and cloned into the eukaryotic expression vector Downstream of the vector pGL3 181hTERT tumor-specific telomerase promoter, a recombinant eukaryotic expression vector pGL3 181hTERT / TRAIL was constructed. The recombinant vector was transfected into human laryngeal carcinoma cell line Hep2 by cationic liposome. The apoptosis of transfected Hep2 cells was observed by trypan blue exclusion method and genomic DNA agarose gel electrophoresis. Results: The recombinant human eukaryotic expression vector pGL3 181hTERT / TRAIL was constructed and its expression product could induce Hep2 laryngeal carcinoma cell apoptosis. CONCLUSION: The recombinant eukaryotic expression vector pGL3 181hTERT / TRAIL has been successfully constructed, which offers the possibility of gene targeting therapy for tumors.