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背景与目的我们的前期研究发现尼古丁能诱导肺癌细胞上皮间质转化。本研究的目的是探讨尼古丁诱导的上皮间质转化(epithelial-mesenchymal transition,EMT)与肺癌侵袭之间的关系。方法应用不同浓度尼古丁处理肺腺癌A549细胞,应用Real-time PCR和Western blot方法检测EMT相关分子标志物E-钙粘蛋白(E-cadherin)和波形蛋白(Vimentin)m RNA和蛋白表达水平,应用免疫荧光技术检测β-链蛋白(β-catenin)蛋白表达位置的变化,应用划痕实验和Transwell小室侵袭实验检测尼古丁对肺癌细胞迁移侵袭能力的影响。结果尼古丁明显下调肺癌细胞株A549 E-cadherin m RNA和蛋白水平表达(P<0.01,P<0.01),并具有浓度和时间依赖性;尼古丁明显上调肺癌细胞株A549 Vimentin m RNA和蛋白水平表达(P<0.01,P<0.01);尼古丁诱导肺癌细胞株A549细胞β-catenin蛋白发生核转移;划痕实验和侵袭实验观察到尼古丁处理的肺癌细胞株A549细胞的迁移和侵袭能力明显增强(P<0.01,P<0.01)。结论尼古丁能够诱导肺癌细胞发生EMT,并且促进肺癌细胞株A549细胞的体外侵袭潜能。
Background and Objective Our previous study found that nicotine induces epithelial-mesenchymal transition in lung cancer cells. The purpose of this study was to investigate the relationship between nicotine-induced epithelial-mesenchymal transition (EMT) and lung cancer invasion. Methods Lung adenocarcinoma A549 cells were treated with different concentrations of nicotine. Real-time PCR and Western blot were used to detect the expressions of E-cadherin and Vimentin mRNA and protein in EMT-related lung cancer cells. The changes of the expression of β-catenin protein were detected by immunofluorescence. The scratch test and Transwell chamber invasion assay were used to detect the effect of nicotine on the migration and invasion of lung cancer cells. Results Nicotine significantly down-regulated the mRNA and protein expression of E-cadherin in A549 cells (P <0.01, P <0.01), and had a concentration-dependent manner. Nicotine significantly up-regulated the expression of Vimentin mRNA and protein in lung cancer A549 P <0.01, P <0.01). Nicotine induced the nuclear translocation of β-catenin protein in A549 cells. Scratch and invasion assays showed that the migration and invasion ability of nicotine-treated lung cancer cell line A549 cells were significantly enhanced (P < 0.01, P <0.01). Conclusion Nicotine can induce EMT in lung cancer cells and promote the invasive potential of lung cancer cell line A549 in vitro.