靶向性重组TRAIL基因诱导人黑色素瘤细胞株A375凋亡的效应

Apoptotic effect of human maligant melanoma cell line A375 induced by tumor-specific recombinant TRAIL gene

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。方法刺激人外周血淋巴细胞增殖,提取总DNA。采用RT-PCR扩增含有IL-2信号肽和TRAIL功能区的融合基因,克隆入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黑色素瘤细胞株A375中,以台盼蓝拒染法和电镜下细胞形态变化,观察转染的A375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,其表达产物能诱导人黑色素瘤细胞株A375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 explore the targeted gene therapy of tumors by using the TRAIL gene in combination with the telomerase promoter. Methods Total RNA was extracted from peripheral blood lymphocytes （PBL） whose proliferation had been stimulated. The TRAIL, gene with an IL-2 signal peptide gene was amplified by RT-PCR and cloned into the vector pGL3-181hTERT downstream of the hTERT promoter to form a eukaryntic expressing vector. A375 cells were transfected by the recombinant vector and apoptosis of the transfected cells was evaluated by rypan-blne exclusion and transmission electron microscopy （TEM）. Restilts We successfully constructed a recombinant eukaryotic expressinn vector for the TRAIL gene. The expressed product significantly induced the apoptosis of the A375 cells. Conclusion The recombinant eukaryotic expression vector pGL3-IL-181hTERT/ TRAIL was suceessfully constructed, which provides the possibility for gene therapy of tumors.