Inhibition of epithelial-mesenchymal transition in A549 cell by transfected Napsin A

作     者：ZHENG Jin-xu GUAN Shu-hong XU Qing LIU Ji-zhu SONG Ping 

作者机构：Department of Respiratory Medicine Affiliated Hospital ofJiangsu University Zhenjiang Jiangsu 212001 China School of Medical Science and Laboratory Medicine JiangsuUniversity Zhenjiang Jiangsu 212013 China 

出 版 物：《Chinese Medical Journal》 (中华医学杂志（英文版）)

年 卷 期：2012年第125卷第15期

页      面：2734-2740页

核心收录：

学科分类：1002[医学-临床医学] 1001[医学-基础医学(可授医学、理学学位)] 100104[医学-病理学与病理生理学] 10[医学] 

基　　金："333" Project of Jiangsu Province(Leading Group Office of Managing the Talents in Jiangsu Province Scientific Research Project of Ministry of Public Health 

主　　题：epithelial-mesenchymal transition, A549 cell, Napsin A transforming growth factor-β1 focal adhesion kinase 

摘      要：Background Epithelial-mesenchymal transition is a cellular process characterized by the loss of cell adhesion, inhibition of E-cadherin expression, and increased cell mobility. Cells without Napsin A are susceptible to transition. Further studies are required to investigate whether this transition can be reversed by restoration of Napsin A. Methods A Napsin A expression vector PLJM1-Napsin A plasmid was constructed and then transfected into the epithelial cell line A549 by lentivirus transfection to obtain A549-PLJM1-Napsin A cell line. Cell proliferation was assayed by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide and cell cycle was measured by flow cytometry. The E-cadherin, type I collagen, and focal adhesion kinase mRNA level was detected by reverse transcription-polymerase chain reaction. The Napsin A, E-cadherin, type I collagen, and focal adhesion kinase protein level in A549 cells was detected by Western blotting. Results Transforming growth factor-β1 induced epithelial-mesenchymal transition in A549 cells, as demonstrated by significant reduction of E-cadherin mRNA and protein levels （P 〈0.01） as well as up-regulation of type I collagen （P 〈0.01）. Transfection of Napsin A in A549 cells can partially block the transforming growth factor-β1-regulated expression of E-cadherin and type I collagen （P 〈0.01）. In addition, transforming growth factor-β1-induced cell proliferation was inhibited by Napsin A （P 〈0.01）. Further study demonstrated that Napsin A caused G0/G1 arrest and inhibited the expression of focal adhesion kinase （P 〈0.01）, a key protein in the integrin signaling pathway, in the in vitro epithelial-mesenchymal transition model. Conclusions Sustained Napsin A expression in A549 cells can inhibit the transforming growth factor-β1-induced epithelial-mesenchymal transition. This may be due to the Napsin A-mediated inhibition of focal adhesion kinase expression and integrin signaling pathway.