A Histone Deacetylase Inhibitor Suppresses Epithelial-Mesenchymal Transition and Attenuates Chemoresistance in Biliary Tract Cancer.
A Histone Deacetylase Inhibitor Suppresses Epithelial-Mesenchymal Transition and Attenuates Chemoresistance in Biliary Tract Cancer.
复制标题
DOI：
10.1371/journal.pone.0145985
复制
发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Mori M
中科院分区：
文献类型：
--
作者：
Sakamoto T;Kobayashi S;Yamada D;Nagano H;Tomokuni A;Tomimaru Y;Noda T;Gotoh K;Asaoka T;Wada H;Kawamoto K;Marubashi S;Eguchi H;Doki Y;Mori M
上皮-间质转化（EMT）参与了恶性肿瘤的侵袭、转移和化疗耐药等特征。在胆道癌（BTC）中，EMT由转化生长因子-β 1（TGF-β1）诱导。EMT是可逆的;因此，可以想象它可能与某些表观遗传变化有关。我们重点关注组蛋白去乙酰化酶（HDAC）抑制剂作为TGF-β1信号转导的调节剂，并研究它们对EMT和化疗耐药性的影响。我们采用了四种BTC细胞系（MzChA-1、吉西他滨耐药MzChA-1、TFK-1和吉西他滨耐药TFK-1），并使用伏立诺他作为HDAC抑制剂。通过qRT-PCR测量上皮标志物（CDH 1）和间充质标志物（CDH 2、波形蛋白、SNAI 1）的相对mRNA表达，以评估与EMT相关的因素。采用MTT法（3-（4，5-Dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide）检测各组细胞的耐药性。此外，NOD/SCID小鼠用于评价伏立诺他的体内作用。在亲本MzChA-1和TFK-1细胞系中，TGF-β1诱导EMT和化学抗性;而伏立诺他抑制TGF-β1诱导的EMT和化学抗性。在高表达TGF-β1的吉西他滨耐药细胞系中，伏立诺他抑制EMT并减弱化疗耐药性。我们发现伏立诺他抑制TGF-β1的信号传导因子SMAD 4的核转位，这是伏立诺他调节EMT的机制之一。我们还发现伏立诺他减弱了SMAD 4与CDH 1相关转录因子SNAI 1、SNAI 2、ZEB 1、ZEB 2和TWIST的结合亲和力。此外，伏立诺他和吉西他滨的联合治疗改善了用吉西他滨抗性MzChA-1细胞异种移植的小鼠的存活时间。总之，伏立诺他通过抑制SMAD 4核转位调节TGF-β1诱导的EMT和化疗耐药性。
Epithelial-mesenchymal transition (EMT) is involved in the characteristics of malignancy, such as invasion, metastasis, and chemoresistance. In biliary tract cancer (BTC), EMT is induced by transforming growth factor-beta 1 (TGF-β1). The EMT is reversible; therefore, it is conceivable that it could be related to some epigenetic changes. We focused on histone deacetylase (HDAC) inhibitors as regulators of TGF-β1 signaling, and investigated their effect on EMT and chemoresistance. We employed four BTC cell lines (MzChA-1, gemcitabine-resistant MzChA-1, TFK-1, and gemcitabine-resistant TFK-1) and used vorinostat as the HDAC inhibitor. The relative mRNA expression of an epithelial marker (CDH1) and mesenchymal markers (CDH2, vimentin, SNAI1) were measured by qRT-PCR to evaluate factors associated with EMT. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was performed to evaluate the chemoresistance of each cell line. In addition, NOD/SCID mice were used to evaluate the effect of vorinostat in vivo. In the parent MzChA-1 and TFK-1 cell lines, TGF-β1 induced EMT and chemoresistance; while vorinostat inhibited the EMT and chemoresistance induced by TGF-β1. In gemcitabine-resistant cell lines that highly expressed TGF-β1, vorinostat inhibited EMT and attenuated chemoresistance. We showed that vorinostat inhibits nuclear translocation of SMAD4 which is a signaling factor of TGF-β1, and this is one of the mechanisms by which vorinostat regulates EMT. We also showed that vorinostat attenuates the binding affinity of SMAD4 to the CDH1-related transcription factors SNAI1, SNAI2, ZEB1, ZEB2, and TWIST. Furthermore, combination therapy with vorinostat and gemcitabine improved survival time in the mice xenografted with gemcitabine resistant MzChA-1 cells. In conclusion, vorinostat regulated TGF-β1-induced EMT and chemoresistance through inhibition of SMAD4 nuclear translocation.