Abstract
The epithelial-mesenchymal transition (EMT) process strongly contributes to cancer metastasis. This study was to investigate the alteration of EMT-related proteins (ZEB1, ZEB2 and S100A4) in cholangiocarcinoma (CCA) tissues. The effect of tumor necrosis factor-α (TNF-α) on the expression of those molecules in CCA cells was investigated. The quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was used to quantify ZEB1, ZEB2 and S100A4 mRNA levels in 50 CCA tissues and related its expression to clinicopathological data. ZEB2 protein immunostaining was investigated in 165 CCA tissues. The effect of TNF-α on EMT-related CCA cell migration was evaluated using qRT-PCR, immunofluorescence and transwell migration assays. ZEB2 and S100A4 mRNA levels were found to be higher in CCA tissues. High levels of S100A4 mRNA and ZEB2 protein were significantly associated with CCA metastasis (P = 0.04 and P = 0.03). Moreover, a trend toward statistical association was found with high levels of both ZEB2 mRNA and protein with shorter survival time (P = 0.10 and P = 0.19). In addition, TNF-α induced CCA cell migration by the induction of transforming growth factor-β (TGF-β) resulting in ZEB2 and S100A4 mRNA and protein activation. These studies demonstrate that TNF-α plays crucial role in the progression of CCA by activating TGF-β signaling and the induction of ZEB2 and S100A4, EMT-related proteins expression.
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