尼古丁诱导肺癌细胞上皮间质转化促进其侵袭转移  

Nicotine Induced Lung Cancer Cells Epithelial-mesenchymal Transition and Promote Its Vitro Invasion Potential

作　　者：侯艳须 李雪冰[1] 潘振华[1] 祖玲玲[1] 范亚光 尤嘉琮[1] 王玉丽[1] 王岷[1] 陈沛锐 沈旺[1] 周清华[1,2] 

机构地区：[1]天津市肺癌转移与肿瘤微环境重点实验室天津市肺癌研究所天津医科大学总医院,天津300052 [2]四川大学华西医院肺癌中心,成都610041

出　　处：《中国肺癌杂志》2016年第4期169-176,共8页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金(No.81572288);教育部高等学校博士点优先发展领域基金(No.20131202130001)资助~~

摘　　要：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 could induce lung cancer cell epithelial-mesenchymal transition （EMT）. The aim of this study is to explore the relationship between nicotine-induced EMT and lung cancer invasion and metastasis. Methods Real-time PCR and Western blot were used to detect the expression changes of EMT-related markers, E-cadherin and Vimentin, in A549 lung cancer cells treated with nicotine;hTe transposition ofβ-catenin protein expression was determined by immunolfuorescence;Scratch test and Transwell invasion assay were used to detect the effects of nicotine on lung cancer cell migration and invasion. Results Nicotine can signiifcantly down-regulate the expressional level of E-cadherin mRNA and protein of A549 cells in a manner of dose and time-dependent （P〈0.01, P〈0.01）;Nicotine can signiifcantly up-regulate the expressional level of Vimentin mRNA and protein of A549 cells in a manner of dose and time-dependent （P〈0.01, P〈0.01）;Immunolfuorescence results showed thatβ-catenin protein was signiifcantly transfered to nucleus;Scratch test and Transwell assay showed that Nicotine could remarkably increase the migration and invasion poten-tial of lung cancer cells （P〈0.01, P〈0.01）. Conclusion Nicotine can induce cancer cells EMT, and promote the invasion and metastasis ability of lung cancer cells.

关 键 词：肺肿瘤 尼古丁 上皮间质转化 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]