EGF and beta1 integrin convergently regulate migration of A431 carcinoma cell through MAP kinase activation.
We found that the convergently epidermal growth factor (EGF)-induced signal and the collagen-induced signal activate mitogen-activated protein kinase (MAPK), which induces migration. We examined the signaling mechanisms of EGF-induced cell migration on collagen using the A431 carcinoma cell. EGF (10 ng/ml) induced migration on collagen, but inhibited proliferation. Using a MAPK cascade inhibitor, PD98059, it was shown that EGF-induced migration on collagen was mediated by MAPK whereas EGF-induced migration on fibronectin and vitronectin was not. PD98059 also showed that activation of MAPK induced by EGF enhanced the adhesiveness of A431 cells to collagen. By Western blotting analysis, the kinetics of MAPK phosphorylation induced by EGF and collagen was examined separately, and convergently. First of all, EGF without collagen caused transient MAPK phosphorylation. Collagen without EGF caused MAPK to be immediately and transiently dephosphorylated, and rephosphorylated followed by sustained hyperphosphorylation. EGF together with collagen caused an immediate, and sustained, hyperphosphorylation. These facts suggest that the transient MAPK dephosphorylation induced by collagen is required for migration in order to maintain an appropriate level of sustained phosphorylation. Furthermore, we found that adhesion of A431 cells to collagen was blocked by the anti-beta1 integrin antibody or by the mixed antibodies composed of anti-alpha1, -alpha2, and -alpha3 antibodies, indicating that collagen-induced MAPK phosphorylation was mediated through alpha1beta1, alpha2beta1, and alpha3beta1 integrins.
Kawahara
E
E
Department of Laboratory Sciences, School of Health Sciences, Faculty of Medicine, Kanazawa University, 5-11-80 Kodatsuno, Kanazawa, 9200942, Japan. kawahara@kenroku.kanazawa-u.ac.jp
Nakada
Natsuko
N
Hikichi
Tetsuro
T
Kobayashi
Jun
J
Nakanishi
Isao
I
D002465:Cell Movement / Q000502:physiology*; D003094:Collagen; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005353:Fibronectins; D006801:Humans; D019012:Integrin beta1 / Q000502:physiology*; D020935:MAP Kinase Signaling System / Q000502:physiology; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology*; D014407:Tumor Cells, Cultured; D019096:Vitronectin
D005353:Fibronectins; D019012:Integrin beta1; D019096:Vitronectin; D004815:Epidermal Growth Factor; D003094:Collagen; D020928:Mitogen-Activated Protein Kinases