信号素3A对肺癌A549细胞上皮-间充质转化的调节作用  

Effect of Semaphorin3A on regulation of epithelial-mesenchynml transition in human lung adenocarcinoma A549 cell strains

作　　者：周海英[1] 吴爱萍 董洪超[1] 何珺[1] 王宝杰[1] 张逊[3] 

机构地区：[1]唐山市工人医院胸外科,063000 [2]华北理工大学病理系,唐山063009 [3]天津市胸科医院胸外科,300070

出　　处：《中华实验外科杂志》2016年第2期407-410,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察外源性信号素3A（SEMA3A）对转化生长因子-β1（TGF-β1）介导的肺癌A549细胞发生上皮-间充质转化（EMT）的调节作用。方法常规培养肺癌A549细胞并分为3组：对照组、TGF—β1诱导组、SEMA3A干预组，观察细胞形态，采用划痕实验检测细胞迁移能力，免疫细胞化学染色检测波形蛋白（Vimentin）的表达，Western blot法检测E-钙黏蛋白（E—cadherin）和Vimentin的蛋白表达水平。结果予以5雌／LTGF—β1诱导48h后，可见A549细胞由原有的铺路石样，转化成纺锤体形或长梭形，细胞间连接消失，细胞间隔变大，呈离散状态。划痕实验结果显示，TGF-β1能够显著促进A549细胞迁移，迁移率为（75．67±4．51）％，是对照组[（33．00±7．00）％]的2．29倍（P〈0．01）；免疫细胞化学染色结果显示，对照组无Vimentin阳性表达，而予以TGF—β1诱导后，可见Vimentin阳性表达，胞质内出现棕黄色丝状排列结构；Western blot结果显示TGF—β1诱导48h后，Vimentin蛋白表达明显上调，是对照组的6．86倍，E—cadherin蛋白表达明显下调，是对照组的5．70％。而予以SEMA3A预处理后，A549细胞基本保持原有的铺路石样状态，细胞迁移能力降低，Vimentin表达下调，E—cadherin表达上调。结论SEMA3A能够通过上调E—cadherin蛋白的表达，抑制Vimentin蛋白的表达，显著抑制TGF—β1介导的A549细胞发生的EMT，同时抑制TGF-β1诱导的细胞迁移能力的提高，从而发挥抗非小细胞肺癌发生、发展的作用。Objective To explore the effect of Semaphorin3A （SEMA3A） on regulation of epithelial- mesenehymal transition （EMT） in human lung adenocarcinoma A549 cell strains. Methods A549 cells were cultured in vitro and divided to 3 groups： （1） control; （2） transforming growth factor - β1 （TGF - β1 ） ; （3） TGF - β1 ＋ SEMA3A. EMT was observed by inverted phase contrast microscope. Cell migration was detected by wound scratch assay. The expression of Vimentin was measured by Immunocytoehemistric staining. And the levels of E - eadherin and Vimentin was detected by Western blotting. Resuits TGF - β1 - treated A549 cells in vitro gradually transformed from elliptic type to a spindle - like morphology with an Vimentin positive muscle wire structure in cytoplasm. The migration rate in 48 h after treatment with TGF - β1 measured by would healing assay was increased 2. 29 fold [ （75.67 ±4. 51 ）% vs. （33.00 ±7.00） % ]. The Vimentin expression in TGF - β1 group increased to 6. 86 fold and expression of E - cadherin decreased by 5.70% compared with control group. Pretreatment with SEMA3A could inhibited all this changes. Conclusion The results suggest that a novel mechanism of action for SEMA3A＇ s beneficial effect in non - small - cell lung carcinoma （NSCLC） , which involves attenuation of EMT induced by TGF - β1 and cell migration.

关 键 词：肺癌 A549细胞 信号素3A 上皮-间充质转化 

分 类 号：R575.2[医药卫生—消化系统]