rL-RVG通过α7烟碱型乙酰胆碱受体诱导胃癌细胞系SGC凋亡的机制  

Mechanism of rL-RVG-induced apoptosis of gastric cancer cell line SGC via alpha 7-nAChR

作　　者：步雪峰[1] 章安伟 尹超云 张烜烽 陈正威 张日婷 严玉兰[3] BU Xue-feng;ZHANG An-wei;YIN Chao-yun;ZHANG Xuan-feng;CHEN Zheng-wei;ZHANG Ri-ting;YAN Yu-lan(Department of General Surgery, the Affiliated People's Hospital of Jiangsu University, Zhenjiang 212002;School of Medicine,Jiangsu University, Zhenjiang 212013;Department of Respiratory Medicine, the Affiliated People's Hospital of Jiangsu University, Zhenjiang 212002, China)

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

出　　处：《基础医学与临床》2019年第5期623-629,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81672999)

摘　　要：目的探讨表达狂犬病毒疫苗糖蛋白的重组新城疫病毒(rL-RVG)通过乙酰胆碱受体诱导胃癌细胞凋亡的可能机制。方法将对数增殖期的胃癌细胞系SGC随机分成rL-RVG、新城疫病毒(NDV)组、PBS组,并同时分别设α7烟碱型乙酰胆碱受体(α7-nAChR)抑制剂(MLA)及激动剂(ACB)预处理组。病毒感染24 h后,CCK8法测定病毒对SGC增殖影响;蛋白印迹检测α7-nAChR、RVG、NDV;凋亡蛋白cleved-caspase-3、BAX/BCL-2;P-ERK、ERK的表达。免疫荧光法测定P-ERK表达。结果随着rL-RVG、NDV病毒液浓度增加,胃癌细胞活力下降,rL-RVG在10^(-2)稀释倍数时抗增殖作用显著(P<0.05);病毒组凋亡相关蛋白cleved-caspase-3、BAX/BCL-2相对表达量较PBS组明显升高,且rL-RVG组较NDV组及PBS组升高明显,乙酰胆碱受体抑制剂预处理后rL-RVG组、NDV组及PBS组凋亡蛋白水平明显增高,胆碱受体激动剂预处理组凋亡蛋白表达水平明显下降(P<0.05);P-ERK在病毒组表达下降,且rL-RVG组较NDV组更低,乙酰胆碱受体抑制剂、激动剂预处理后分别下降、上升(P<0.05)。结论rL-RVG通过α7-nAChR诱导胃癌细胞系SGC凋亡的机制之一是抑制ERK通路。Objective To explore the possible mechanism of the recombinant avirulent NDV LaSota strain expressing the rabies virus glycoprotein (rL-RVG) inducing the apoptosis of gastric cancer cells by acetylcholine receptor pathway. Methods The gastric cancer cell line SGC at logarithmic proliferation stage were randomly divided into the rL-RVG , Newcastle disease virus(NDV)and PBS groups.Furthermore,the acetylcholine receptor(α7-nAChR)inhibitor groups and the agonist pretreatment groups were set up respectively.At 24 hours after infected by virus, CCK8 assay was used to determine the inhibitory effect of virus on the growth of gastric cancer cell lines. Western blot detected the protein expression of alpha 7-nAChR, RVG, NDV and apoptosis-related proteins including cleved-caspase-3 and BAX/BCL-2 as well as the expression of P-ERK and ERK. Immunofluorescence was used to examine the expression of P-ERK. Results The concentrations of the recombinant and wild virus inhibition of growth of gastric cancer cell line SGC were greater than 10 -2 /mL and 10 -3 /mL, and the rL-RVG group was significantly more inhibited than NDV and PBS group( P <0.05).Western blot showed apoptosis-related protein cleved-caspase-3, BAX/BCL-2 in rL-RVG group and NDV group increased significantly as compared to PBS group, the protein expression of P-ERK/ERK in the virus group was lower than the control group. The apoptosis-related protein expression levels in the rL-RVG/NDV/PBS groups pretreated with choline receptor inhibitor significantly increased than that in the groups without pretreatment.Besides,the groups pretreated with choline receptor agonist had decreased apoptosis protein expression as compared with the groups without pretreating.Western blot and immunofluorescence showed the expression of P-ERK decreased in the virus group, and the recombinant virus group decreased significantly compared with the NDV and PBS group ( P < 0.05).Furthermore, the P-ERK protein expression in groups pretreated with acetylcholine receptor inhibitor descended.On

关 键 词：重组新城疫病毒 狂犬疫苗糖蛋白 胃癌 烟碱型乙酰胆碱受体 ERK 

分 类 号：R735.2[医药卫生—肿瘤]