Evidence of TGF-ß1 mediated epithelial-mesenchymal transition in immortalized benign prostatic hyperplasia cells.
Mol Membr Biol
; 31(2-3): 103-10, 2014.
Article
em En
| MEDLINE
| ID: mdl-24650126
ABSTRACT
Expression of epithelial-mesenchymal transition (EMT) markers has been detected clinically in benign prostatic hyperplasia (BPH) tissues. To understand the molecular basis, we investigated the role of stromal microenvironment in the progression of EMT in BPH cells. First, we used cell culture supernatant from normal prostate stromal WPMY-1 cells to provide supernatant-conditioned medium (WSCM) to culture the BPH-1 cell line. Then, the morphological changes and migratory capacity were detected in BPH-1 cells. The expression of EMT markers was examined in BPH-1 cells by Western blot and immunofluorescent analysis. Finally, to investigate the role of transforming growth factor beta 1 (TGF-ß1) in this process, the WSCM-cultured cells were treated with monoclonal antibody against TGF-ß1 to study its effect on EMT. We found that the morphology of BPH-1 cells changed to a spindle-like shape after cultured in WSCM, and the levels of E-cadherin and cytokeratin 5/8 (CK5/8) were significantly lower than the cells cultured in ordinary medium. These BPH-1 cells were also tested positive for mesenchymal markers vimentin and a-smooth muscle actin (SMA) as well as Snail. We also found WSCM can increase the migratory capacity of BPH-1 cells. In addition, when they were treated with anti-TGF-ß1, upregulation of E-cadherin and CK5/8 levels was observed but no expression of vimentin, alpha-SMA or Snail was detected. Furthermore, phosphorylated-Smad3 expression in WSCM-cultured BPH-1 cells was also suppressed by anti-TGF-ß1 treatment. Our results demonstrated that stromal cell supernatant was able to induce EMT in BPH-1 cells, possibly through secreting TGF-ß1 to activate Smad signaling. Our results suggest novel molecular targets for clinical treatment of BPH by modification of stromal microenvironment through inhibiting TGF-ß1/Smad expression.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Hiperplasia Prostática
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Transdução de Sinais
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Células Estromais
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Meios de Cultivo Condicionados
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Fator de Crescimento Transformador beta1
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Transição Epitelial-Mesenquimal
Limite:
Humans
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Male
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article