Related Experiment Video
Updated: Nov 1, 2025
Induction and Analysis of Epithelial to Mesenchymal Transition
Published on: August 27, 2013
Trichostatin A promotes esophageal squamous cell carcinoma cell migration and EMT through BRD4/ERK1/2-dependent
Danhui Liu1,2, Yuzhen Liu1,2,3, Bo Qi1,2
1Department of Thoracic Surgery, The First Affiliated Hospital of Xinxiang Medical University, Weihui, China.
Background:
Histone deacetylases (HDACs) have been demonstrated to be aberrantly activated in tumorigenesis and cancer development. Thus, HDAC inhibitors (HDACIs) are considered to be promising anti-cancer therapeutics. However, recent studies have shown that HDACIs promote the migration of many cancer cells. Therefore, there is a need to elucidate the underlying mechanisms of HDACIs on cancer cell migration to establish a combination therapy that overcomes HDACI-induced cell migration.
Methods:
KYSE-150 and EC9706 cells were treated differently. Effects of drugs and siRNA treatment on tumor cell migration and cell signaling pathways were investigated by transwell migration assy. Gene expression for SNAI2 was tested by RT-qPCR. Western blot analysis was employed to detect the level of E-cadherin, β-catenin, vimentin,Slug，ERK1/2, H3, PAI-1 and BRD4. The effect of drugs on cell morphology was evaluated through phase-contrast microscopic images.
Results:
TSA promotes epithelial-mesenchymal transition (EMT) in ESCC cells by downregulating the epithelial marker E-cadherin and upregulating mesenchymal markers β-catenin, vimentin, Slug, and PAI-1. Knockdown of Slug by siRNA or inhibition of PAI-1 clearly suppressed TSA-induced ESCC cell migration and resulted in the reversal of TSA-triggered E-cadherin, β-catenin, and vimentin expression. However, no crosstalk between Slug and PAI-1 was observed in TSA-treated ESCC cells. Blocking ERK1/2 activation also inhibited TSA-induced ESCC cell migration, EMT, and upregulation of Slug and PAI-1 levels in ESCC cells. Interestingly, inhibition of BRD4 suppressed TSA-induced ESCC cell migration and attenuated TSA-induced ERK1/2 activation and upregulation of Slug and PAI-1 levels.
Conclusions:
Our data indicate the existence of at least two separable ERK1/2-dependent signaling pathways in TSA-mediated ESCC cell migration: an ERK1/2-Slug branch and an ERK1/2-PAI-1 branch. Both branches of TSA-induced ESCC cell migration appear to favor the EMT process, while BRD4 is responsible for two separable ERK1/2-dependent signaling pathways in TSA-mediated ESCC cell migration.
Insights
Histone deacetylase inhibitors (HDACIs) like TSA can promote cancer cell migration by inducing epithelial-mesenchymal transition (EMT). Targeting ERK1/2 and BRD4 pathways may offer strategies to overcome HDACI-induced cell migration in esophageal squamous cell carcinoma (ESCC).
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Aberrant activation of histone deacetylases (HDACs) is implicated in tumorigenesis.
- HDAC inhibitors (HDACIs) show anti-cancer potential but can paradoxically promote cancer cell migration.
- Understanding HDACI mechanisms in cancer cell migration is crucial for developing effective combination therapies.
Purpose of the Study:
- To elucidate the mechanisms by which HDAC inhibitors promote cancer cell migration.
- To investigate the role of specific signaling pathways, including ERK1/2, Slug, PAI-1, and BRD4, in HDACI-induced migration.
- To identify potential therapeutic targets for overcoming HDACI-induced cell migration in esophageal squamous cell carcinoma (ESCC).
Main Methods:
- Treatment of ESCC cell lines (KYSE-150 and EC9706) with Trichostatin A (TSA).
- Assessment of cell migration using transwell assays.
- Analysis of gene and protein expression (E-cadherin, β-catenin, vimentin, Slug, ERK1/2, PAI-1, BRD4) via RT-qPCR and Western blotting.
- Evaluation of cell morphology and pathway inhibition using siRNA and specific inhibitors.
Main Results:
- TSA treatment induced epithelial-mesenchymal transition (EMT) in ESCC cells, characterized by decreased E-cadherin and increased β-catenin, vimentin, Slug, and PAI-1.
- Knockdown of Slug or inhibition of PAI-1 suppressed TSA-induced migration and reversed EMT markers.
- Inhibition of ERK1/2 activation and BRD4 suppressed TSA-induced ESCC cell migration, EMT, and the upregulation of Slug and PAI-1.
Conclusions:
- TSA-mediated ESCC cell migration involves at least two distinct ERK1/2-dependent pathways: one involving Slug and another involving PAI-1.
- Both pathways promote EMT, contributing to TSA-induced cell migration.
- BRD4 plays a critical role in regulating these separable ERK1/2-dependent signaling pathways in TSA-mediated ESCC cell migration.
Related Concept Videos
02:38Mitogens and the Cell Cycle
01:12Role Of Notch Signalling In Intestinal Stem Cell Renewal
Direct cell-to-cell contact is needed for the activation of Notch signaling. The signal is initiated when a notch ligand binds to a receptor on an adjacent cell, also...
03:03mTOR Signaling and Cancer Progression
The mTOR pathway or the...
01:22PI3K/mTOR/AKT Signaling Pathway
01:21Barrett Esophagus-I: Introduction
This constant acid exposure transforms the esophagus's pink mucosal lining (stratified squamous epithelium) into a type of lining more...
02:30Metastasis
Epithelial-to-Mesenchymal Transition
The epithelial-to-mesenchymal transition or EMT is a developmental process commonly observed in wound healing, embryogenesis, and cancer metastasis. EMT is induced by transforming growth factor-beta (TGF-β) or receptor tyrosine kinase (RTK) ligands, which further...