狂犬病毒糖蛋白重组的新城疫病毒诱导人胃癌SGC细胞自噬

The rabies virus glycoprotein recombinated Newcastle disease virus induce the human stomachal adenocarcinoma autophagy

目的:观察狂犬病毒糖蛋白重组的新城疫病毒(rabies virus glycoprotein recombinated Newcastle disease virus,r L-RVG)及野生型新城疫病毒(Newcastle disease virus,NDV)对胃癌SGC细胞自噬的影响。方法:将SGC细胞随机分为3组,rl-RVG组(转染rl-RVG)、NDV组(转染NDV)、对照组(转染PBS),24 h后观察细胞的形态变化,评价病毒对细胞的影响。用免疫双标记和免疫印迹法鉴定病毒感染是否成功;蛋白质印迹检测各组细胞自噬相关蛋白LC3及Beclin1的表达;透射电镜观察细胞内超微结构的变化和自噬体的形成。结果:病毒感染细胞24 h后,rl-RVG组及NDV组细胞皱缩明显,且rl-RVG组较NDV组变化明显,对照组无明显变化;rl-RVG组和NDV组细胞LC3及Beclin1蛋白表达较对照组明显升高(P<0.05),且rl-RVG组明显高于NDV组(P<0.05);rl-RVG组及NDV组出现的自噬体明显多于对照组,且rl-RVG组较NDV组明显增多(P<0.05)。结论:r L-RVG较NDV更易引起胃癌SGC细胞的自噬。

Objective:To observe the autophagy of the human gastric cancer SGC infected by rabies virus glycoprotein recombinated Newcastle disease virus(rL-RVG)or wild type Newcastle disease virus (NDV).Methods:The cells were divided into three groups at random,the rL-RVG group(infected with rL-RVG),the NDV group(infected with NDV),and the control group(infected with PBS).The mor-phology of SGC cells after infected 24 hours were monitored by microscope.The infected cells were ana-lysed with immunofluorescence,the total proteins get from the cells infected with or without virus were analysed with western blotting,and the ultrastructural of SGC was observed by transmission electron mi-croscope.Results:After infected 24 hours,there was a significantly morphologic changes in the rL-RVG group and the NDV group,and the cells were shrink,the rL-RVG group changes were more markedly than the NDV group,and the control group had no change;compared with the control group,the expression of the autophagy associated proteins levels in rL-RVG group and NDV group were higher(P <0.05),and the rL-RVG group showed higher levels than the NDV group(P <0.05).There was more autophage bodies showed in the rL-RVG group and the NDV group,and the rL-RVG group had even more autophage bodies than the NDV group(P <0.05).Conclusion:The rL-RVG could introduce more autophagy than the NDV.