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
首发时间:2008-10-27
摘要: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48h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。
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Construction of Eukaryotic Expression Vector of NT4-APOPTIN-HA2-TAT in secretory form and its effects on the tumor cells
Abstract:Objective: To explore antitumor effect of Eukaryotic Expression Vector of NT4-APOPTIN-HA2-TAT in secretory form. Methods: Recombination vector fusing with Apoptin(VP3 )、NT4、HA2-TAT were constructed and were transfected to HepG2 and NIH3T3 cells.The results of cell death were detected by MTT and Flowcytometre assay. Results:Recombined adeno-associated virus fusing NT4-Apoptin-HA2 -TAT could induce apoptosis after HepG2 cells were transfected in 48h later,but not in normal NIH3T3 cells. Conclusion: NT4-APOPTIN-HA2-TAT can be secreted and induces apoptosis of tumor cells specificity,and it provides a new way of anti-tumor gene therapy.
Keywords: secretory apoptin eukaryotic expression vector apoptosis of tumor cells
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