Bombesin stimulates invasion and migration of Isreco1 colon carcinoma cells in a Rho-dependent manner.
The membrane receptor for the neuropeptide bombesin/gastrin-releasing peptide (GRP) is expressed by a large fraction of human colorectal carcinoma cells. We reported previously a stimulation of cell adhesion and lamellipodia formation by the neuropeptide bombesin in the human, bombesin/GRP receptor-expressing, Isreco1 colorectal cancer cell line (J. C. Saurin et al., Cancer Res., 59: 962-967, 1999). Using invasion and motility assays, we demonstrate in this report that bombesin can both enhance the invasive capacity of Isreco1 cells in a dose-dependent manner (maximal effect at 1 nM) and stimulate the closure of wounds performed on confluent Isreco1 cells. These effects were reversed fully by the specific bombesin/GRP receptor antagonist D-Phe(6)-Bn(6-13)OMe used at 1 micro M. MMP-9 and urokinase-type plasminogen activator were expressed by Isreco1 cells, and bombesin did not significantly alter their level of secretion. Interestingly, exoenzyme C3 (10 micro g/ml) decreased cell invasiveness induced by bombesin by 70% and completely inhibited the migration of Isreco1 cells. Similarly, the Rho-kinase inhibitor Y-27632 dose-dependently reduced the effect of bombesin on cell invasion. Moreover, pull-down assays for GTP-bound RhoA demonstrated that bombesin was able to activate the small G-protein in Isreco1 cells. These results show that the neuropeptide bombesin is able to modulate invasiveness of Isreco1 colorectal carcinoma cells in vitro through a Rho-dependent pathway, leading to an increase in cell locomotion without a significant effect on tumor-cell associated proteolytic activity. These findings indicate that bombesin/GRP receptor expression may contribute to the cellular events that are critical for invasion/migration of colorectal carcinoma cells.
INSERM Unité 45 and IFR 62, Hôpital Edouard-Herriot, Pavillon H, F-69437 Lyon Cedex 3, France. saurin@lyon151.inserm.fr
Jean-Christophe
JC
Saurin
Marjorie
M
Fallavier
Bernard
B
Sordat
Jean-Claude
JC
Gevrey
Jean-Alain
JA
Chayvialle
Jacques
J
Abello
D001839:Bombesin / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects* / Q000502:physiology; D003094:Collagen; D003110:Colonic Neoplasms / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D004338:Drug Combinations; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007797:Laminin; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009361:Neoplasm Invasiveness; D017346:Protein Serine-Threonine Kinases / Q000502:physiology*; D011509:Proteoglycans; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000502:physiology; D020741:rho GTP-Binding Proteins / Q000502:physiology*; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein / Q000502:physiology
D004338:Drug Combinations; D047908:Intracellular Signaling Peptides and Proteins; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; D014568:Urokinase-Type Plasminogen Activator; D020780:Matrix Metalloproteinase 9; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein; D001839:Bombesin