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目的:探讨上皮间质转化(epithelial-mesenchymal transition,EMT)过程在肺癌侵袭转移中的作用。方法:体外培养A549细胞,以bFGF(10ng/ml)进行干预后,倒置相差显微镜观察细胞形态学变化;间接免疫荧光观察上皮细胞标志物E-cadherin和间质细胞标志物vimentin蛋白表达的变化;采用细胞划痕试验检测bFGF对A549细胞迁移能力的影响;采用transwell小室试验检测bFGF对A549细胞侵袭能力的影响。结果:bFGF(10ng/ml)干预后,在倒置相差显微镜下观察,A549细胞形态变成了梭形,形态如同成纤维细胞。间接免疫荧光显示A549细胞E-cadherin表达随时间延长逐渐减弱,而vimentin表达逐渐增强。细胞划痕试验显示,bFGF干预后细胞迁移能力提高。Transwell小室试验显示,bFGF干预后细胞侵袭能力提高。结论:bFGF在体外诱导肺腺癌细胞系A549细胞发生上皮间质转化,上皮间质转化是肺癌侵袭转移的重要机制之一。
Objective: To investigate the role of epithelial-mesenchymal transition (EMT) in invasion and metastasis of lung cancer. Methods: A549 cells were cultured in vitro and the morphological changes of cells were observed by inverted phase contrast microscope after intervention with bFGF (10ng / ml). The expression of E-cadherin and vimentin in epithelial cells was detected by indirect immunofluorescence staining. The cell scratch assay was used to detect the effect of bFGF on the migration of A549 cells. The transwell assay was used to detect the effect of bFGF on the invasiveness of A549 cells. Results: After treated with bFGF (10ng / ml), the morphology of A549 cells turned into a fusiform shape and observed as an inverted phase contrast microscope. The morphology of the cells was similar to that of fibroblasts. Indirect immunofluorescence showed that the expression of E-cadherin in A549 cells gradually decreased with time and the expression of vimentin gradually increased. Cell scratch test showed that bFGF increased cell migration ability after intervention. Transwell chamber assay showed that cell invasion ability increased after bFGF intervention. Conclusion: bFGF induces epithelial-mesenchymal transition in A549 cells and epithelial-mesenchymal transition is an important mechanism of lung cancer invasion and metastasis.