目的探讨细胞内钙离子络合剂1,2-二(2-氨基苯氧基)乙烷-N,N,N',N'-四乙酸四乙酰氧甲基酯[1,2-bis(2-aminophenoxy)ethane-N,N,N'N'-tetraacetic acid,BAPTA-AM]对转化生长因子(transformation growth factor-β,TGF-β)诱导人肝癌细胞Hep G2上皮-间叶化转变(epithelium-mesenchymal transition,EMT)的影响。方法 TGF-β、TGF-β联合BAPTA-AM分别作用Hep G2细胞24 h后,采用Real time-PCR方法检测上皮细胞钙粘素(E-cadherin)mRNA的表达变化,采用Transwell方法检测Hep G2细胞迁移及侵袭的能力。结果与对照组比较,TGF-β处理Hep G2细胞24 h后,细胞形态变梭,E-cadherin mRNA表达下调(P〈0.05),细胞的迁移、侵袭能力均增加(P〈0.01);与TGF-β组比较,TGF-β联合BAPTA-AM处理Hep G2细胞24 h后,E-cadherin mRNA表达上调(P〈0.05),细胞的迁移、侵袭能力均减弱(P〈0.01)。结论 TGF-β能诱导肝癌细胞Hep G2发生EMT,增强Hep G2的迁移、侵袭能力,BAPTA-AM能抑制TGF-β所诱导Hep G2细胞发生EMT,其机制可能与E-cadherin mRNA的表达上调有关。
Objective To explore the effects of intracellular calcium ion chelating agent[ 1,2 -bis (2 -amino- phenoxy) ethane - N, N, N" N'- tetraacetic acid ] ( BAPTA - AM ) on transforming growth factor - β ( TGF - β ) induced the epithelium - mesenchymal transition (EMT) of human hepatocellular carcinoma HepG2 cells. Methods The mRNA expression of E - cadherin in HepG2 cells after treated with TGF - β alone and combined with BAPTA - AM for 24 h were analyzed by Real time - PCR, the migration and invasion ability of HepG2 cells were estimated in transwell assays. Results Compared with control group, the morphology of HepG2 cells were changed spindle, and the E - cadherin mRNA level of HepG2 ceils were decreased significantly after TGF - β treatment for 24 h (P 〈 0.05 ), which were increased after TGF - β combined with BAPTA - AM treatment for 24 h (P 〈 0.05 ) ; however the migration and invasion capability of HepG2 cells were enhanced distinctly (P 〈 0.01 ) after TGF - β treatment for 24 h, which were obviously weakened ( P 〈 0.01 ) after TGF - β combined with BAPTA - AM treatment for 24 h. Conclusion TGF - β could induce the EMT of HepG2 cells and promote the migration and invasion ability of HepG2 cell, which were inhibited by BAPTA - AM, and the mechanism may be related with the upregulation of E - cadherin mRNA expression.