rL-RVG体外抑制肺癌细胞增殖并促进其凋亡  

RL-RVG inhibits proliferation and promotes apoptosis of lung cancer cells in vitro

作　　者：严玉兰[1] 梁冰[1,2] 张金[1] 贾丽娟[1,2] 刘洋[1] 张杰[2] 

机构地区：[1]江苏大学附属人民医院呼吸科,江苏镇江212002 [2]江苏大学临床医学院,江苏镇江212001

出　　处：《细胞与分子免疫学杂志》2014年第5期449-452,457,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：江苏省镇江市社会发展项目(2013041)

摘　　要：目的将表达狂犬病毒糖蛋白的重组LaSota株新城疫病毒疫苗(rL-RVG)感染至A549肺腺癌细胞,观察其对肺癌细胞增殖和细胞凋亡的影响。方法采用直接感染的方法将rL-RVG感染至A549细胞,采用Western blot法检测rL-RVG中RVG和NDV蛋白在A549细胞内的表达;MTT法检测rL-RVG对细胞增殖的影响,TUNEL法检测rL-RVG对A549细胞凋亡的影响;AnnexinⅤ-FITC/PI染色结合流式细胞检测术检测A549细胞凋亡情况;Western blot法检测caspase-3的表达,并与对照组LaSota株进行比较,PBS组为空白对照组。结果 A549细胞感染rL-RVG后RVG蛋白和NDV蛋白都稳定表达,MTT法结果显示细胞增殖明显被抑制,rL-RVG组抑制率高于LaSota组。A549细胞凋亡增多,其中流式细胞检测术中显示rL-RVG组早期凋亡细胞较其他2组增多,差异有统计学意义(P<0.05),TUNEL检测凋亡指数增加,rL-RVG组较其他2组增多,差异有统计学意义(P<0.05)。Western blot法显示促凋亡蛋白caspase-3表达增加,加入caspase抑制剂Z-VAD-FMK后,caspase-3蛋白表达明显降低。结论 rL-RVG感染A549细胞后能在细胞内稳定表达,rL-RVG可抑制肺癌细胞生长、促进肺癌细胞凋亡,效果优于野生LaSota株。Objective To infect lung adenocarcinoma A549 cells with recombinant LaSota strain Newcastle disease virus （NDV） vaccine expressing the rabies virus glycoprotein （rL-RVG）, and explore the effect of rL-RVG on proliferation and apoptosis of lung cancer cells. Methods A549 cells were infected with the rL-RVG and then detected for the expressions of RVG and NDV proteins by Western blotting. The cell proliferation was examined by Ml-r assay and apoptosis index and cell early apoptosis were respectively detected by TUNEL and annexin V-FITC/PI staining combined with flow cytometry. The expression of pro-apoptotic protein caspase-3 was observed by Western blotting. The LaSota strain of NDV was used as control group and PBS was the blank control. Results Both RVG and NDV proteins were stably expressed in A549 cells infected with rL-RVG. Ml-r assay results showed that cell proliferation was significantly inhibited, and the inhibition rate of the rL-RVG group was higher than that of LaSota group. The apoptosis of A549 cells were promoted by rL-RVG infection. Flow cytometry revealed that the early apoptotic cells of the rL-RVG group increased as compared with the other two groups （P〈0.05）. Consistently, TUNEL assay showed the apoptotic index increased in the rL-RVG group as compared with the other two groups （ P 〈 0.05）. Western blotting demonstrated that the expression of the pro-apoptotic protein caspase-3 was up-regulated. However, when we added specific broad-spectrum caspase inhibitor Z-VAD-FMK, the expression of the caspase-3 protein was obviously reduced. Conclusion The rL-RVG is stably expressed in the infected A549 cells. The rL-RVG inhibits lung cancer cell growth and promote cell apoptosis, and the effect of rL-RVG is better than the wild LaSota strain.

关 键 词：新城疫病毒 基因重组 肺癌细胞 凋亡 细胞增殖 

分 类 号：R734.2[医药卫生—肿瘤] Q279[医药卫生—临床医学]