摘要
目的观察表达狂犬病毒糖蛋白的重组新城疫病毒(rl-RVG)对人肺癌细胞系A549的迁移影响,初步探讨其可能的机制。方法重组新城疫病毒rl-RVG直接感染A549细胞为rl-RVG实验组,新城疫病毒La Sota株处理的A549细胞组及未感染病毒的A549细胞组作为对照组。Western blot法检测NDV-HN蛋白及狂犬病毒糖蛋白(RVG),细胞增殖实验测定新城疫病毒使用的最佳作用浓度;划痕实验及Transwell法测A549迁移;Western blot法及免疫荧光法检测E-cadherin,MMP2蛋白表达。结果空白对照组无NDV、rl-RVG表达,rl-RVG仅在感染rl-RVG细胞组有表达,NDV在感染rl-RVG细胞组及感染NDV细胞组皆有表达;与空白对照组相比,rl-RVG及NDV稀释浓度大于5×10-5对细胞的增殖有抑制作用(P<0.05);rl-RVG组迁移的距离及细胞数明显减少(P<0.05)。与空白对照组及NDV感染组相比,rl-RVG组E-cadherin蛋白表达水平上调(P<0.05),MMP2蛋白表达水平减弱(P<0.05)。结论重组新城疫病毒rl-RVG能抑制人肺癌细胞系A549的迁移,并可能通过影响肺腺癌A549上皮细胞-间质转化(EMT)过程中的调控因子E-cadherin,MMP2蛋白而对细胞迁移而作用。
Objective Observe the effects of rl-RVG on migration of lung adenocarcinoma A549cells, preliminarily explore potential mechanism. Methods The group infected with the rl-RVG was experimental group, The group infected with NDV and the group not infected with virous were control groups. The experimental detected for the ex- pressions of RVG and NDV-HN proteins by Western blot. Cell growth experiment determined the best active concen- trations of newcastle disease virus and rl-RVG; Observe the effects of rl-RVG and NDV on migration of lung adeno- carcinoma A549 cells by scratch assay and Transwell method. The expression of E-cadherin and MMP2 was ob- served by Western blot and immunofluorescence. The LaSota strain of NDV was used as control group and PBS was the blank control. Results Neither RVG nor NDV proteins didn＇t expressed in blank control group. RVG proteinexpressed in rl-RVG group and NDV protein expressed inhibited in rl-RVG group and NDV group more signifi in both rl-RVG group and NDV group. Cell proliferation was cantly as compared with the blank control group（P 〈.0. 05 ）. After infected with rl-RVG, migration distance and number of cells significantly reduced （ P 〈 0. 05 ）. After A549 cells were infected with rl-RVG, the expression of E-cadherin was enhanced （P 〈 0. 05 ）and the expression of MMP2 was decreased as compared with the blank control group and NDV group （ P 〈 0. 05 ）. Conclusions Recom- binant avirulent newcastle disease virus can inhibit the migration of A549 cells in vitro, which may be attributed to regulatory factors of E-eadherin and MMP2 in the procession of epithelial mesenchymal transition EMT.
出处
《基础医学与临床》
CSCD
2015年第4期508-513,共6页
Basic and Clinical Medicine
基金
镇江市社会发展基金(2013041)
