摘要
目的探讨表达鸡新城疫病毒(NDV)血凝素-神经氨酸酶(HN)基因的重组腺病毒Ad-HN对人肝癌细胞HepG-2的抑制作用。方法应用Ad-HN感染HepG-2细胞,采用MTT法检测Ad-HN对HepG-2细胞增殖的抑制作用;AnnexinV染色法结合流式细胞仪检测细胞凋亡;AO/EB染色法和DAPI染色法对Ad-HN感染的肿瘤细胞及细胞核进行形态学观察;底物显色法检测Caspase1、Caspase3、Caspase6和Caspase8活性的变化。结果Ad-HN能够有效抑制HepG-2细胞的增殖,当感染剂量为100MOI,作用时间为48h时,Ad-HN对HepG-2细胞增殖的抑制率达峰值(17.20%～35.04%);AnnexinV染色结果显示,Ad-HN感染48h后HepG-2细胞凋亡率为29.39%;Ad-HN感染导致HepG-2细胞呈现细胞浓缩、细胞核皱缩进而碎裂等凋亡特征;Ad-HN感染可显著上调HepG-2细胞的Caspase酶活性。结论Ad-HN能够诱导HepG-2细胞凋亡,并对HepG-2细胞产生抑制效应。
Objective To investigate the inhibitory effect of recombinant adenovirus Ad-HN expressing hemagglutinin（HA）neuramidinase（NA）gene of Newcastle disease virus （NDV）on human hepatoma HepG-2 cells. Methods HepG-2 cells were infected with Ad-HN, and the inhibitory effect of Ad-HN on proliferation of HepG-2 cells was determined by MTT method. The apoptosis of HepG-2 cells was determined by Annexin V staining combined with flow cytometry. The morphologies of HepG-2 cells infected with Ad-HN and their nuclei were observed by AO / EB staining and DAPI staining. The activities of Caspase 1, 3, 6 and 8 were determined by substrate coloration. Results Ad-HN inhibited the proliferation of HepG-2 cells effectively. After infection with Ad-HN at a MOI of 100 for 48 h, the inhibition rate of HepG-2 cells reached a peak value of 17. 20% ～ 35. 04%. Annexin V staining showed that the apoptosis rate of HepG-2 cells 48 h after infection with Ad-HN was 29. 39%. The infection with Ad-HN induced the apoptotic character of HepG-2 cells, such as cell condensation as well as nuclear shrinkage and breakage. Meanwhile, the infection with Ad-HN significantly up-regulated the Caspase activity of HepG-2 cells. Conclusion Ad-HN induced the apoptosis of and showed inhibitory effect on HepG-2 cells.
出处
《中国生物制品学杂志》
CAS
CSCD
2010年第4期350-353,共4页
Chinese Journal of Biologicals
基金
国家高技术发展计划(863计划
2007AA021004)
关键词
新城疫病毒
血凝素-神经氨酸酶基因
重组腺病毒
肝癌
抗肿瘤作用
Newcastle disease virus（NDV）
Hemagglutinin（HA）-neuramidinase（NA）gene
Recombinant adenovirus
Hepatoma
Antitumor effect
