SMAD3 is essential for transforming growth factor-β1-induced urokinase type plasminogen activator expression and migration in transformed keratinocytes

Abstract

Transforming growth factor-β1 (TGF-β1) stimulates the extracellular matrix degrading proteases expression and cell migration in order to enhance cancer cells malignancy. In the present study, we analysed the role of TGF-β1-induced Smad3 activation in the urokinase type plasminogen activator (uPA) production, as well as in cell migration and E-cadherin downregulation in transformed PDV keratinocyte cell line. TGF-β1 signalling was interfered by the chemical inhibitor of the TGF-β1-receptor 1 (ALK5), SB505124, and the specific Smad3 inhibitor, SiS3. Our results showed that TGF-β1 stimulates uPA expression directly through ALK5 activation. The inhibition of Smad3 strongly reduced the capacity of TGF-β1 to stimulate uPA expression, in parallel decreasing the uPA inhibitor plasminogen activator inhibitor type 1 (PAI-1) expression. In addition, the transient expression of dominant negative Smad3 mutant inhibited the TGF-β1-induced uPA promoter transactivation. Moreover, Smad3-/- mouse embryonic fibroblasts were refractory to the induction of uPA by TGF-β1. The inhibition of both ALK5 and Smad3 dramatically blocked the TGF-β1-stimulated E-cadherin downregulation, F-actin reorganisation and migration of PDV cells. Taken together, our results suggest that the TGF-β1-induced activation of Smad3 is the critical step for the uPA upregulation and E-cadherin downregulation, which are the key events preceding the induction of cell migration by TGF-β1 in transformed cells.

Ets Related Gene and Smad3 Proteins Collaborate to Activate Transforming Growth Factor-Beta Mediated Signaling Pathway in ETS Related Gene-Positive Prostate Cancer Cells

“…The TGF- β signaling pathway have a dual role in cancer progression, which is a tumor suppressor for normal epithelial and early stages of cancer cells, and also, it is a tumor promoter in final stages of the metastatic diseases (Kocic, 2012, Miles, 2012, Tarasewicz, 2012). Our results provide support that ERG plays a major role in prostate cancer progression through the regulation of TGF- β signaling pathway.…”

“…Phosphoryated Smd2 and Smad3 interact with Smad4 and shown translocate into the nucleus as a complex where they bind to transcriptional co-factors and regulate transcription of target genes (Feng, 2005, Massague, 2012, Sakaki-Yumoto, 2013). TGF- β signaling pathway has been accepted to have a dual role in tumor progression, which is a tumor suppressor for normal epithelial and early stages of cancer cells, and the other is a tumor promoter in last steps of the metastatic disease (Kocic, 2012, Miles, 2012). …”

Interleukin-17 modulates myoblast cell migration by inhibiting urokinase type plasminogen activator expression through p38 mitogen-activated protein kinase

“…For F-actin staining, phalloidin coupled to TRITC (Sigma-Aldrich) was used (Kocić et al, 2012b). Images were taken with a microscope equipped with epi-fluorescence…”

Diabetes-Induced Renal Injury in Rats Is Attenuated by Suramin

“…In addition, the profibrotic adhesion molecule OPN was shown to enhance glomerular damage in DN mouse models, probably through the TGF-␤ 1 pathway, whereas deletion of OPN protected against progression of DN (Nicholas et al, 2010). Whereas the mechanism by which suramin reverses the appearance of all these proteins in STZ rat urine is not known, it is possible that it is the result of suramin blocking TGF-␤ 1 signaling (Nicholas et al, 2010;Kocic et al, 2012). Thus, the application of LC-MS/MS techniques may provide crucial insights into tubular and glomerular changes that cannot be revealed by using traditional renal functional and pathological parameters and also highlight potential pathogenic mechanisms not previously known or considered for DN.…”