title: Promotion of Metastasis-associated Gene Expression in Survived PANC-1 Cells Following Trichostatin A Treatment.
abstract: OBJECTIVE
  Histone deacetylase inhibitors represent a promising class of potential anticancer agents for the treatment of human malignancies. In this study, the effects of trichostatin A (TSA) on apoptosis, metastasis-associated gene expression, and activation of the Notch pathway in human pancreatic cancer cell lines were investigated.
  
  
  METHODS
  After treatment with TSA, cell viability and apoptosis were evaluated using the MTT [3-(4,5-dimethylthia-zol-2-yl)-2,5-diphenyltetrazolium bromide] assay, Hoechst 33258 staining, and flow cytometry. Moreover, RT-PCR and western blot analyses were performed to measure the expression levels of apoptosis-associated genes (Bcl-2, Bax, and caspase-3), metastasis-associated genes (E-cadherin, vimentin, and matrix metalloproteinases), and Notch pathway activation (Notch intracellular domain, NICD). The levels of matrix metalloproteinase 2 and NICD were also semi-quantified by immunoassay.
  
  
  RESULTS
  Following treatment with TSA for 24 h, PANC-1, SW1990, and MIATACA-2 cells exhibited cell death. The MTT assay revealed that TSA significantly decreased cell viability in a dose-dependent manner in PANC-1 cells. The Hoechst 33258 staining and flow cytometry results evidenced a significant increase in PANC-1 cell apoptosis following TSA treatment. The expression levels of Bax and caspase-3 were increased significantly, whereas Bcl-2 was down-regulated after TSA treatment. In the PANC-1 cells that survived after TSA treatment, the expression levels of vimentin, E-cadherin, and MMP genes were altered by the promotion of potential metastasis and increased expression of NICD.
  
  
  CONCLUSIONS
  TSA can induce apoptosis of pancreatic cancer cells. In addition, the up-regulation of metastasis-related genes and the activation of the Notch pathway in the survived PANC-1 cells may be associated with a too-low level of TSA or resistance to TSA.
authors.forename: Zongjing
authors.initials: Z
authors.lastname: Chen
authors.forename: Yunxiu
authors.initials: Y
authors.lastname: Yang
authors.forename: Biao
authors.initials: B
authors.lastname: Liu
authors.forename: Benquan
authors.initials: B
authors.lastname: Wang
authors.forename: Meng
authors.initials: M
authors.lastname: Sun
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
authors.forename: Bicheng
authors.initials: B
authors.lastname: Chen
authors.affiliation: Department of Surgery, the First Affiliated Hospital of Wenzhou Medical University, 2 Fuxue Lane, Wenzhou 325000, Zhejiang, P.R. China. bisonch@163.com.
authors.forename: Heyi
authors.initials: H
authors.lastname: You
authors.forename: Mengtao
authors.initials: M
authors.lastname: Zhou
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D015870:Gene Expression / Q000187:drug effects*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D009362:Neoplasm Metastasis / Q000235:genetics*
chemical_list: D006877:Hydroxamic Acids; C012589:trichostatin A