摘要
目的本实验研究旨在探讨含新城疫病毒HN基因的重组质粒对人胃癌细胞BGC-823凋亡的联合作用机制。方法将含有HN基因的重组质粒pIRVP3IL-18HN经脂质体介导转染人胃癌细胞BGC-823,通过MTT方法检测细胞活力;电镜下观察细胞形态;罗丹明123和DCFA染色测定线粒体跨膜电位(△Ψ)和活性氧(ROS)水平变化;底物染色反应检测Caspase-3活性。结果重组质粒pIRVP3IL-18HN转染人胃癌细胞BGC-823后,MTT法结果显示致肿瘤细胞死亡率升高,细胞活力下降;电镜观察肿瘤细胞呈现明显凋亡状态;与阴性空白质粒对照显示,线粒体△Ψ下降,ROS水平升高;Caspase-3活性被激活。结论含新城疫病毒HN基因的重组质粒可以诱导人胃癌细胞BGC-823进入特定的凋亡程序,从而导致人胃癌细胞BGC-823的凋亡。
Objective To study the mechanism of apoptosis in Human Gastric carcinoma cells BGC-823 Induced with recombinant plasmid comprised Hemagglutinin-neuraminidase（HN） Gene from Newcastle Disease Virus（NDV）.Methods The recombinant plasrnid pIRVP3IL-18HN was introduced into human gastric carcinoma cells BGC-823 by liposome-mediated transfeetion.rlhe viability of BGC-823 cells was determined by MTT staining and the morphology of BGC-823 was observed by electron microscope. The alteration of mitochondrial transmembrane potential and ROS level of the cells was detected by flow cytometry （FCM） with Rhodamine 123 and DCFA staining and the activation of caspase-3 was assayed by substrate color reaction. Results After recombinant plasmid was introduced into BGC-823 cells, mortality of cells was increased and viability was decreased significantly respectively. Apoptosis of BGC-823 cells can be observed significantly by electron microscope. Compared with control cells, mitochondrial transmembrane potential was decreased, ROS level of the cells was increased, and caspase-3 was activated. Conclusion A sequerming apoptosis of human gastric carcinoma cells BGC-823 can be induced with recombinant plasmid comprised Hemngglutinin-rmuraminidase （HN） Gene from Newcastle Disease Virus,then induce the apoptosis of tumor cells.
出处
《中国实验诊断学》
2006年第5期441-445,共5页
Chinese Journal of Laboratory Diagnosis
基金
国家"973"资助项目(No.G1999011902)
关键词
新城疫病毒
删基因
重组质粒
人胃癌细胞BGC-823
凋亡
newcastle disease virus
HN gene
recombinant plasmid
human gastric carcinoma cells BGC-823, apoptosis
