重组新城疫病毒rL-RVG诱导人胃腺癌细胞内质网应激和凋亡  

Recombinant Newcastle disease virus( rL-RVG) triggers apoptosis by inducing endoplasmic reticulum stress in gastric carcinoma cell

作　　者：赵英海[1,2] 严玉兰[3] 李米[1,2] 王穆彬[1] 步雪峰[2] 

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]江苏大学附属人民医院普外科,江苏镇江212002 [3]江苏大学附属人民医院呼吸科,江苏镇江212002

出　　处：《江苏大学学报（医学版）》2016年第3期199-203,共5页Journal of Jiangsu University：Medicine Edition

基　　金：江苏省自然科学基金资助项目(BK20151333)

摘　　要：目的:观察新城疫病毒(newcastle disease virus,NDV)及稳定表达狂犬病毒糖蛋白的重组新城疫病毒(recombinant avirulent NDV La Sota strain expressing the rabies virus glycoprotein,rL-RVG)对胃腺癌HGC细胞内质网应激及凋亡的影响。方法:将HGC细胞随机分为对照组、NDV组和rL-RVG组,分别转染PBS、NDV和rL-RVG 24 h,免疫印迹法和免疫荧光检测糖调节蛋白78(glucose-regulated protein 78,GRP78)和CCAAT/增强子结合蛋白同源蛋白(CCAAT/enhancer-binding protein homologous protein,CHOP)表达。再将HGC细胞随机分为对照组、4-苯基丁酸钠盐(sodium phenylbutyrate,4-PBA)组、rL-RVG组和4-PBA+rL-RVG组,4-PBA组和4-PBA+rL-RVG组分别加入4-PBA预处理4 h,然后rL-RVG组和4-PBA+rL-RVG组分别转染rL-RVG,对照组和4-PBA组转染PBS,免疫印迹法检测各组细胞GRP78、CHOP和半胱氨酸蛋白酶-3(caspase-3)的表达,流式细胞术检测细胞凋亡率。结果:感染HGC细胞24 h后,NDV组及rL-RVG组GRP78和CHOP表达明显高于对照组(P均<0.05),且rL-RVG组明显高于NDV组(P<0.01)。4-PBA预处理后,4-PBA+rL-RVG组GRP78、CHOP、caspase-3蛋白表达和细胞凋亡率较rLRVG组明显降低(P<0.05)。结论:rL-RVG较NDV更易引起胃腺癌HGC细胞内质网应激,且与凋亡相关。Objective： To investigate the effects of newcastle disease virus （NDV） and recombinant avirulent NDV LaSota strain expressing the rabies virus glycoprotein （rL-RVG） on endoplasmic reticulum stress and apoptosis of HGC cells. Methods： The cells were divided into three groups at random, control group, NDV group, and rL-RVG group, which infected with PBS, NDV and rL-RVG, respectively. The expression of endoplasmic reticulum stress related proteins, glucose-regulated protein 78 （ GRP78 ）, CCAAT/enhancer-binding protein homologous protein（CHOP） were analyzed by western blotting and im- munofluorescence. The HGC cells were divided into four groups at random, control group, 4-PBA group, rL-RVG group and 4-PBA ＋ rL-RVG group. 4-PBA group and 4-PBA ＋ rL-RVG group were treated with sodium phenylbutyrate（4-PBA） for 4 h, then rL-RVG group and 4-PBA ＋ rL-RVG group were infected with rL-RVG, while rL-RVG were replaced with PBS in control group and 4-PBA group. The expression of caspase-3 protein was detected with western blotting. The rate of cell early apoptosis were detected by flow cytometry. Results： After infected 24 hours, compared with the control group, NDV group and rL- RVG group had high levels of GRP78 and CHOP protein expression （P 〈 0.05 ） , and rL-RVG groupshowed higher levels than NDV group（P 〈0.01 ）. The expression of GRP78, CHOP, caspase-3 protein and the rate of apoptosis in 4-PBA ＋ rL-RVG group was lower than rL-RVG group after pretreated with 4- PBA （P 〈 0.05 ）. Conclusion： The rL-RVG could induce stronger endoplasmic reticulum stress and re- lated apoptosis than the NDV.

关 键 词：新城疫病毒 狂犬病毒糖蛋白 内质网应激 细胞凋亡 

分 类 号：R393[医药卫生—基础医学]