TGF-β Signal Transduction in Pancreatic Carcinoma Cells is Sensitive to Inhibition by the Src Tyrosine Kinase Inhibitor AZM475271.
BACKGROUND
Earlier results from our group have shown that in pancreatic ductal adenocarcinoma (PDAC)-derived cells transforming growth factor (TGF)-β1-dependent epithelial-mesenchymal transition (EMT) and cell motility was inhibited by the Src inhibitors PP2 and PP1 both of which targeted the TGF-β receptors for inhibition.


OBJECTIVE
In this study we evaluated the impact of another Src inhibitor, AZM475271, on various TGF-β responses in PDAC cells.


METHODS
The effect of AZM475271 on TGF-β1-induced random cell migration (chemokinesis), the expression of EMT and migration/invasion-associated genes, TGF-β-induced luciferase activity, and C-terminal phosphorylation of Smad2 and Smad3 was measured in the PDAC-derived Panc-1 and Colo357 cell lines using real-time cell migration assays, quantitative real-time PCR, luciferase reporter gene assays and phosphoimmunoblotting, respectively.


RESULTS
AZM475271 effectively blocked TGF-β1-induced chemokinesis of Panc-1 cells in a dose-dependent fashion and inhibited the high chemokinetic activity of Panc-1 cells with ectopic expression of a constitutively active ALK5T204D mutant. AZM475271 but not another Src inhibitor, SU6656, partially relieved the suppressive effect of TGF-β1 on E-cadherin and inhibited TGF-β1-induced upregulation of the MMP2, MMP9, N-cadherin and vimentin genes, activity of a TGF-β1-dependent reporter gene, and activation of Smad2 and Smad3.


CONCLUSIONS
Our data suggest that AZM475271 cross-inhibits tumor-promoting TGF-β signaling and may thus function as an inhibitor of both TGF-β and Src in both experimental and clinical therapies against metastatic dissemination in late-stage PDAC.
First Department of Medicine, UKSH, Campus Lübeck, 23538 Lubeck. Germany.
Tobias
T
Bartscht
First Department of Medicine, UKSH, Campus Lübeck, 23538 Lubeck. Germany.
Benjamin
B
Rosien
Department of Radiation Oncology, UKSH, Campus Lubeck, D-23538 Lubeck. Germany.
Dirk
D
Rades
Department of General, Visceral and Vascular Surgery, Jena University Hospital, D- 07747 Jena. Germany.
Roland
R
Kaufmann
First Department of Medicine, UKSH, Campus Lubeck, 23538 Lubeck,. Germany.
Harald
H
Biersack
First Department of Medicine, UKSH, Campus Lubeck, 23538 Lubeck,. Germany.
Hendrik
H
Lehnert
First Department of Medicine, University of Lubeck, 23538 Lubeck. Germany.
Hendrik
H
Ungefroren
D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D006801:Humans; D010179:Pancreas / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D010190:Pancreatic Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D010879:Piperazines / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D019061:src-Family Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
C547126:AZM551248; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D016212:Transforming Growth Factor beta; D019061:src-Family Kinases
ALK5; AZM475271; PDAC; Smad; TGF-&#946;; chemokinesis