HDAC inhibitors induce epithelial-mesenchymal transition in colon carcinoma cells
HDAC inhibitors induce epithelial-mesenchymal transition in colon carcinoma cells
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DOI：
10.3892/or.2015.3879
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发表时间：
2015-05-01
期刊：
影响因子：
4.2
通讯作者：
Park, Seon Mee
中科院分区：
文献类型：
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作者：
Ji, Meiying;Lee, Eun Jeoung;Park, Seon Mee
组蛋白去乙酰化酶（HDAC）抑制剂对上皮-间质转化（EMT）的影响在各种类型的癌症中不同。我们研究了四种结肠癌细胞系在用HDAC抑制剂阿司他丁A（TSA）和丙戊酸（VPA）攻击时，有或没有转化生长因子-β 1（TGF-β 1）处理的EMT表型。使用了四种在致瘤性、微卫星稳定性和DNA突变方面具有不同表型的结肠癌细胞系。在用TSA（100或200 nM）或VPA（0.5 mM）加或不加TGF-β 1（5 ng/ml）处理24 h后，使用蛋白质印迹分析、免疫荧光、定量实时RT-PCR通过E-钙粘蛋白和波形蛋白的表达来评估EMT表型。还通过细胞形态学、迁移和侵袭测定来评估生物EMT表型。TSA或VPA通过在mRNA和蛋白水平上减少E-钙粘蛋白和增加波形蛋白表达诱导结肠癌细胞中的间充质特征。共聚焦显微镜显示E-钙粘蛋白的膜衰减或核移位以及波形蛋白的表达增强。这些反应发生在6 h后，并增加至24 h。TSA或VPA处理后，结肠癌细胞由圆形或矩形变为梭形，迁移和侵袭能力增强。在微卫星稳定细胞（SW 480和HT 29）和微卫星不稳定细胞（DLD 1和HCT 116）中，TSA或VPA诱导的EMT变化的敏感性相当。TSA或VPA在结肠癌细胞中诱导间充质表型，并且这些作用在TGF-β 1存在下增强。HDAC抑制剂在作为结肠癌的新抗癌药物应用之前需要谨慎。
The effects of histone deacetylase (HDAC) inhibitors on epithelial-mesenchymal transition (EMT) differ in various types of cancers. We investigated the EMT phenotype in four colon cancer cell lines when challenged with HDAC inhibitors trichostatin A (TSA) and valproic acid (VPA) with or without transforming growth factor-beta 1 (TGF-beta 1) treatment. Four colon cancer cell lines with different phenotypes in regards to tumorigenicity, microsatellite stability and DNA mutation were used. EMT phenotypes were assessed by the expression of E-cadherin and vimentin using western blot analysis, immunofluorescence, quantitative real-time RT-PCR following treatment with TSA (100 or 200 nM) or VPA (0.5 mM) with or without TGF-beta 1 (5 ng/ml) for 24 h. Biological EMT phenotypes were also evaluated by cell morphology, migration and invasion assays. TSA or VPA induced mesenchymal features in the colon carcinoma cells by a decrease in E-cadherin and an increase in vimentin expression at the mRNA and protein levels. Confocal microscopy revealed membranous attenuation or nuclear translocation of E-cadherin and enhanced expression of vimentin. These responses occurred after 6 h and increased until 24 h. Colon cancer cells changed from a round or rectangular shape to a spindle shape with increased migration and invasion ability following TSA or VPA treatment. The susceptibility to EMT changes induced by TSA or VPA was comparable in microsatellite stable (SW480 and HT29) and microsatellite unstable cells (DLD1 and HCT116). TSA or VPA induced a mesenchymal phenotype in the colon carcinoma cells and these effects were augmented in the presence of TGF-beta 1. HDAC inhibitors require careful caution before their application as new anticancer drugs for colon cancers.