论文部分内容阅读
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 Ad-HN, a recombinant adenovirus expressing NDV hemagglutinin-neuraminidase (HN) gene on human hepatoma HepG-2 cells. Methods Ad-HN was used to infect HepG-2 cells. MTT assay was used to detect the inhibitory effect of Ad-HN on HepG-2 cell proliferation. Annexin V staining and flow cytometry were used to detect apoptosis. AO / EB staining and DAPI staining The morphology of tumor cells and nuclei infected with Ad-HN was observed. The changes of Caspase1, Caspase3, Caspase6 and Caspase8 activities were detected by substrate colorimetric assay. Results Ad-HN inhibited the proliferation of HepG-2 cells at a dose of 100 MOI for 48 h, and the inhibitory rate of Ad-HN on HepG-2 cells reached the peak (17.20% -35.04%). The results of Annexin V staining The results showed that the apoptotic rate of HepG-2 cells was 29.39% at 48h after Ad-HN infection. The apoptosis of HepG-2 cells induced by Ad-HN infection resulted in cell apoptosis and cell ruffling, Upregulate Caspase activity in HepG-2 cells. Conclusion Ad-HN can induce apoptosis in HepG-2 cells and inhibit HepG-2 cells.