Thrombin stimulates integrin β1-dependent adhesion of human pancreatic cancer cells to vitronectin through protease-activated receptor (PAR)-1.
OBJECTIVE
The aim of this study was to investigate the effect of thrombin and the thrombin receptor protease-activated receptor (PAR)-1 on adhesion of human pancreatic cancer cell lines to extracellular matrices (ECMs) and to identify related integrins with these effects.


METHODS
Human pancreatic cancer cell lines SUIT-2 and its four sublines, and Panc- 1, AsPC-1 and MiaPaCa-2 were treated with thrombin, PAR-1 agonist TRAP-6, PAR-1 antagonist SCH79797, or anti-integrin ±vβ3, ±vβ5 and β1 monoclonal antibodies. Cells were incubated for 45 minutes on micro titer plates that were pre-coated with ECMs (fibronectin, laminin, vitronectin, type IV collagen). The number of adherent cells was measured by the MTT method.


RESULTS
Eight human pancreatic cancer cell lines expressed PAR-1. Thrombin significantly enhanced adhesion of SUIT-2 and its sublines and MiaPaCa-2 to vitronectin, especially in the SUIT-2 subline S2-007. We obtained similar results on S2-007 cells through treatment with TRAP-6. However, SCH79797 inhibited the effect of thrombin. Furthermore, anti-integrin β1 antibody conspicuously inhibited 1U/mL thrombin-induced enhancement of adhesion to vitronectin.


CONCLUSIONS
Thrombin significantly enhanced adhesion of pancreatic cancer cells to vitronectin through PAR- 1 depending on the presence of integrin β1. Suppression of thrombin action by anti-integrin β1 antibody will become a useful therapy against pancreatic cancer.
Department of Surgical Oncology and Regulation of Organ Function, Miyazaki University School of Medicine, Miyazaki, Japan.
Mikio
M
Kanemaru
Naoki
N
Maehara
Takeshi
T
Iwamura
Kazuo
K
Chijiiwa
D000230:Adenocarcinoma / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D005109:Extracellular Matrix / Q000502:physiology; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000276:immunology / Q000378:metabolism; D019012:Integrin beta1 / Q000276:immunology / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D010446:Peptide Fragments / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D044463:Receptor, PAR-1 / Q000378:metabolism*; D019030:Receptors, Vitronectin / Q000276:immunology / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology*; D019096:Vitronectin / Q000502:physiology
D000911:Antibodies, Monoclonal; D039302:Integrin alphaVbeta3; D019012:Integrin beta1; C415424:N3-cyclopropyl-7-((4-(1-methylethyl)phenyl)methyl)-7H-pyrrolo(3, 2-f)quinazoline-1,3-diamine; D010446:Peptide Fragments; D011758:Pyrroles; D011799:Quinazolines; D012333:RNA, Messenger; D044463:Receptor, PAR-1; D019030:Receptors, Vitronectin; D019096:Vitronectin; C064660:integrin alphaVbeta5; C082835:thrombin receptor peptide (42-47); D013917:Thrombin