基因-病毒治疗系统CNHK200-hA对肺癌治疗的研究

CNHK200-hA-a gene viral therapeutic system and its antitumor effect on lung cancer

目的 构建抗肿瘤新生血管生成的基因治疗与病毒治疗相结合的基因 病毒治疗系统CNHK2 0 0 hA ,并对其肺癌的体内外治疗作用进行初步研究。方法 通过Westernblot分析 ,观察携带抗肿瘤新生血管生成基因angiostatink1 5 (hA)的基因 病毒治疗系统CNHK2 0 0 hA和非增殖型腺病毒对介导k1 5蛋白表达的影响 ;通过细胞病理作用及动物试验 ,比较CNHK2 0 0 hA和非增殖型腺病毒对人肺癌细胞株A5 4 9的杀伤作用及对肺癌裸鼠移植瘤的治疗作用。结果 基因 病毒治疗系统CNHK2 0 0 hA能够介导hA基因在肺癌细胞内高效表达k1 5蛋白 ,其表达量高于非增殖型腺病毒Ad hA。细胞病理效应显示 ,CNHK2 0 0 hA对A5 4 9的杀伤作用明显优于Ad hA ,CNHK2 0 0 hA在感染复数 (MOI)值为 1时可完全杀伤A5 4 9细胞 ,而达到相同杀伤效应的Ad hA的MOI值为 1 0 0。动物试验显示 ,CNHK2 0 0 hA对肺癌的疗效明显好于Ad hA及肿瘤增殖型病毒ONYX 0 1 5。结论 插入了抗肿瘤新生血管生成基因hA的基因 病毒治疗系统CNHK2 0 0 hA ,可明显提高hA基因的表达量 ,其在体内外的抗肺癌作用较非增殖型腺病毒Ad hA及肿瘤增殖型病毒ONYX 0 1 5进一步提高。

Objective To develop a novel vector system, which combines the advantages of the gene therapy, antiangiogenic therapy and virus therapy, and to observe its effect on lung cancer. Methods Human angiostatin gene hA(k1 5) was inserted into the genome of the replicative virus specific for the tumor cells by virus recombination technology. The expression of hA(k1 5), its effect on tumor growth in vitro and in vivo were studied. Results A new kind of gene viral vector system, designated as CNHK200 hA(k1 5), in which the E1b55 000 gene was deleted but the E1a gene of adenovirus preserved, was constructed. The novel vector system possessed the same property as the replicative virus ONYX 015, which replicates in p53 tumor cells but not in normal cells, thus specifically kills tumor cells. In vitro, CNHK200 hA and Ad hA both could kill A549 tumor cells but the latter needed 100 times more MOI to achieve the same amplitude of cell killing. In vivo, the therapeutic effect of CNHK200 hA on human lung cancer A549 xenograft in nude mice was significantly better than that of Ad hA and that of tumor replicative virus ONYX 015. Conclusion CNHK200 hA (k1 5), a novel vector is constructed in which the angiostatin gene is inserted into the genome of the replicative adenovirus cytotoxic to p53 negative tumor cells. It has the advantages of specific tumor targeting, high level gene expression in tumor cells, and potent tumoricidal activity.