分泌型凋亡素真核表达载体的构建及对肝癌细胞HepG2凋亡的影响

目的 构建含有NT4和HA2-TAT的分泌型VP3真核表达载体,并观察其表达对人肝癌细胞HepG2和小鼠成纤维细胞NIH3T3的影响.方法 构建了含鸡贫血病毒VP3基因和NT4、HA2-TAT基因的分泌型真核表达载体--NT4-Apoptin-HA2-TAT重组腺相关病毒载体.体外转染人肝癌细胞HepG2和小鼠成纤维细胞NIH3T3,用MTT法和流式细胞学,检测凋亡素融合基因诱导细胞凋亡的效果.结果 携带融合基因的重组腺相关病毒感染人肝癌HepG2细胞48 h后,显示了很强的诱导肿瘤细胞凋亡的能力.而对小鼠成纤维细胞NIH3T3细胞无明显凋亡作用.结论 分泌型NT4-Apoptin-HA2-TAT重组腺相关病毒载体转染细胞后能够被正常分泌、穿膜并特异性诱导肿瘤细胞凋亡,为肿瘤的基因治疗提供了新的思路.

Construction of eukaryotic expression vector of NT4-APOPTIN-HA2-TAT in secretory form and its effects on the tumor cells

Objective To explore antitumor effect of eukaryotic expression vector of NT4-Apoptin-HA2-TAT in secretory form.Methods Recombinant vector fusing with Apoptin (VP3), NT4, HA2-TAT were constructed and transfected into HepG2 and NIH3T3 cells.The results of cell death were detected by MTT and flow cytometry.Results The recombined adeno-associated virus fusing NT4-Apoptin-HA2-TAT could induce apoptosis 48h after HepG2 cells were transfected.However, it was not seen in normal NIH3T3 cells.Conclusion NT4-Apoptin-HA2-TAT can be secreted to specifically induce apoptosis of tumor cells, which provides a new way of anti-tumor gene therapy.