摘要目的 探讨真核细胞翻译起始因子5A-2(eIF5A-2)的表达在非小细胞肺癌(NSCLC)上皮间质化(EMT)过程中的作用及其机制.方法 通过Westrn-blotting和免疫荧光检测细胞表型变化,利用siRNA转染下调eIF5A-2表达后再以Western-blotting和免疫荧光检测细胞表型的改变.用TGF-β1刺激诱导NSCLC细胞系NCI-H358和HCC827发生EMT后并利用Western-blotting检测eIF5A-2的表达.结果 NCI-H358和HCC827细胞系低表达eIF5A-2,为上皮表型,而NCI-H1299细胞高表达eIF5A-2,为间质表型.eIF5A-2 siRNA干扰后可以改变NCI-H1299细胞的间质表型.NCI-H358和HCC827细胞经TGF-β1诱导后可发生EMT变化,而eIF5A-2 siRNA干扰后则可防止这种变化的发生.结论 在不同的NSCLC细胞系中,间质表型细胞较上皮表型者高表达eIF5A-2.TGF-β1可诱导上皮表型细胞发生EMT,而干扰eIF5A-2则可阻止EMT的发生.

abstractsObjective To investigate the effect and mechanism of eIF5A-2 in regulating epithelial-mesenchymal transition(EMT) in non-small cell lung cancer (NSCLC).Methods Western-blotting and immunofluorescence have been used to tested phenotype of different NSCLC cell lines.After interfering with eIF5A-2 siRNA,we used western-blotting and immunofluorescence to test phenotype changes.TGF-β1 was used to induce NCI-H358 and HCC827 cells underwent EMT changes.Results NCI-H358 and HCC827 expressed low levels of eIF5A-2 and belonged to epithelial phenotype; while NCI-H1299 expressed high level of eIF5A-2 and belonged to mesenchymal phenotype.After down-regulating eIF5A-2 level by interfering with eIF5A-2 siRNA,NCI-H1299 changed to epithelial phenotype.NCI-H358 and HCC827 changed to mesenchymal phenotypes after stimulating with TGF-β1 and eIF5A-2 siRNA could prevent this transition.Conclusion Mesenchymal phenotype NSCLC cells express high level of eIF5A-2.TGF-β1 could induce epithelial phenotype NSCLC cells undergo EMT changes and interfering eIF5A-2 could prevent this transition.