Background:Epithelial-mesenchymal transition(EMT) is believed to be the critical process in malignant tumor invasion and metastases,and has a great influence on improving the survival rate in non-small-cell lung cance...Background:Epithelial-mesenchymal transition(EMT) is believed to be the critical process in malignant tumor invasion and metastases,and has a great influence on improving the survival rate in non-small-cell lung cancer(NSCLC) patients.Recent studies suggested that eukaryotic initiation factor 5A-2(eIF5A-2) might serve as an adverse prognostic marker of survival.We detected eIF5A-2 in NSCLC A549 cells,and found that the invasive capability correlates with the eIF5A-2 expression.Methods:Transforming growth factor(TGF)-β1 was used to induce EMT in A549 cells.Western blotting,immunofluorescence,wound healing assay,and transwell-matrigel invasion chambers were used to identify phenotype changes.Western blotting was also used to observe changes of the expression of eIF5A-2.We down-regulated the eIF5A-2 expression using an eIF5A-2 siRNA and identified the phenotype changes by western blotting and immunofluorescence.We tested the change of migration and invasion capabilities of A549 cells by the wound healing assay and transwell-matrigel invasion chambers.Results:After stimulating with TGF-β1,almost all A549 cells changed to the mesenchymal phenotype and acquired more migration and invasion capabilities.These cells also had higher eIF5A-2 protein expression.Down-regulation of eIF5A-2 expression with eIF5A-2 siRNA transfection could change the cells from mesenchymal to epithelial phenotype and decrease tumor cell migration and invasive capabilities significantly.Conclusions:The expression of eIF5A-2 was up-regulated following EMT phenotype changes in A549 cells,which correlated with enhanced tumor invasion and metastatic capabilities.Furthermore,in the A549 cell line,the process of EMT phenotype change could be reversed by eIF5A-2 siRNA,with a consequent weakening of both invasive and metastatic capabilities.展开更多
目的:探究钙/钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)在肺腺癌组织中的表达及其促进肺腺癌的侵袭、转移。方法:通过免疫组织化学染色(immunohistochemistry,IHC)分析肺腺癌患者的石蜡组织标本...目的:探究钙/钙调蛋白依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)在肺腺癌组织中的表达及其促进肺腺癌的侵袭、转移。方法:通过免疫组织化学染色(immunohistochemistry,IHC)分析肺腺癌患者的石蜡组织标本CaMKⅡ表达与临床病理参数关系,同时对肺腺癌组织与其配对淋巴结转移病灶中的CaMKⅡ表达情况进行比较分析,收集2011年1月至2011年12月于天津医科大学肿瘤医院行手术治疗的113例肺腺癌患者及21例配对的原发病灶及淋巴结转移病灶的石蜡组织标本。将肺腺癌细胞系H1299及Calu3进行慢病毒转染,实验组转染高表达CaMKⅡ病毒,对照组转染阴性对照病毒,通过Transwell实验和划痕实验检测肺腺癌细胞侵袭、转移能力,通过Western blot检测CaMKⅡ表达水平与上皮间充质转化(pithelial-mesenchymal transition,EMT)相关指标和表皮生长因子受体(epidermal growth factor receptor,EGFR)通路激活间的关系。结果:CaMKⅡ高表达与肺腺癌TNM分期和淋巴结转移呈正相关,肺腺癌淋巴结转移病灶中的癌组织CaMKⅡ表达明显高于其原发病灶(P<0.05)。细胞实验表明,CaMKⅡ高表达的肺腺癌细胞,其穿膜细胞数目明显增多,伤口愈合能力增强;EGFR通路激活后p-CaMKⅡ水平增加,且CaMKⅡ促进了EMT相关蛋白及转录因子的表达。结论:CaMKⅡ参与EGFR信号传导,促进EMT过程,增加肺腺癌的侵袭转移能力。展开更多
基金Project supported by the Natural Science Fundation of Ningbo (No. 2011A610052)the Zhejiang Provincial Natural Science Fundation (No. LY12H16002) of China
文摘Background:Epithelial-mesenchymal transition(EMT) is believed to be the critical process in malignant tumor invasion and metastases,and has a great influence on improving the survival rate in non-small-cell lung cancer(NSCLC) patients.Recent studies suggested that eukaryotic initiation factor 5A-2(eIF5A-2) might serve as an adverse prognostic marker of survival.We detected eIF5A-2 in NSCLC A549 cells,and found that the invasive capability correlates with the eIF5A-2 expression.Methods:Transforming growth factor(TGF)-β1 was used to induce EMT in A549 cells.Western blotting,immunofluorescence,wound healing assay,and transwell-matrigel invasion chambers were used to identify phenotype changes.Western blotting was also used to observe changes of the expression of eIF5A-2.We down-regulated the eIF5A-2 expression using an eIF5A-2 siRNA and identified the phenotype changes by western blotting and immunofluorescence.We tested the change of migration and invasion capabilities of A549 cells by the wound healing assay and transwell-matrigel invasion chambers.Results:After stimulating with TGF-β1,almost all A549 cells changed to the mesenchymal phenotype and acquired more migration and invasion capabilities.These cells also had higher eIF5A-2 protein expression.Down-regulation of eIF5A-2 expression with eIF5A-2 siRNA transfection could change the cells from mesenchymal to epithelial phenotype and decrease tumor cell migration and invasive capabilities significantly.Conclusions:The expression of eIF5A-2 was up-regulated following EMT phenotype changes in A549 cells,which correlated with enhanced tumor invasion and metastatic capabilities.Furthermore,in the A549 cell line,the process of EMT phenotype change could be reversed by eIF5A-2 siRNA,with a consequent weakening of both invasive and metastatic capabilities.
