title: Cellular fibronectin 1 promotes VEGF-C expression, lymphangiogenesis and lymph node metastasis associated with human oral squamous cell carcinoma.
abstract: Lymph node metastasis (LNM) is associated with poor survival in patients with oral squamous cell carcinoma (OSCC). Vascular endothelial growth factor-C (VEGF-C) is thought to be responsible for increased lymphangiogenesis and LNM. Understanding of the mechanism by which VEGF-C expression is regulated in OSCC is thus important to design logic therapeutic interventions. We showed that inoculation of the SAS human OSCC cells expressing the venus GFP (V-SAS cells) into the tongue in nude mice developed LNM. V-SAS cells in LNM were isolated by FACS and re-inoculated into the tongue. This procedure was repeated eight times, establishing V-SAS-LM8 cells. Differential metastasis PCR array between the parental V-SAS and V-SAS-LM8 was performed to identify a molecule responsible for lymphangiogenesis and LNM. Fibronectin 1 (FN1) expression was elevated in V-SAS-LM8 cells compared to V-SAS-cells. V-SAS-LM8 tongue tumor showed increased expression of FN1 and VEGF-C, and promoted lymphangiogenesis and LNM compared with V-SAS tumor. Further, phosphorylation of focal adhesion kinase (FAK), a main downstream signaling molecule of FN1, was up-regulated, and epithelial-mesenchymal transition (EMT) was promoted in V-SAS-LM8 cells. Silencing of FN1 by shRNA in V-SAS-LM8 cells decreased FAK phosphorylation, VEGF-C expression and inhibited lymphangiogenesis and LNM. EMT was also reversed. The FAK phosphorylation inhibitor PF573228 also decreased VEGF-C expression and reversed EMT in V-SAS-LM8 cells. Finally, we detected intense FN1 expression in some clinical specimens obtained from OSCC patients with LNM. These results demonstrate that elevated expression of cellular FN1 and following activation of FAK lead to increased VEGF-C expression, lymphangiogenesis and LNM and promoted EMT in SAS human OSCC cells and suggest that FN1-phosphorylated FAK signaling cascade is a potential therapeutic target in the treatment of LNM in OSCC.
authors: Departments of Molecular and Cellular Biochemistry, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan.|Oral and Maxillofacial Surgery II, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan.|Department of Oral and Maxillofacial Surgery, Seichokai Hannan Municipal Hospital, Hannan, Osaka, Japan.
authors: Yoshihiro
authors: Y
authors: Morita
authors: Departments of Molecular and Cellular Biochemistry, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan.
authors: Kenji
authors: K
authors: Hata
authors: Departments of Molecular and Cellular Biochemistry, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan.
authors: Masako
authors: M
authors: Nakanishi
authors: Department of Oral and Maxillofacial Surgery, Kinan Hospital, Tanabe, Wakayama, Japan.
authors: Tetsuji
authors: T
authors: Omata
authors: Department of Oral and Maxillofacial Surgery, Seichokai Hannan Municipal Hospital, Hannan, Osaka, Japan.|Department of Oral and Maxillofacial Surgery, Kinan Hospital, Tanabe, Wakayama, Japan.|Department of Oral-Maxillo-facial Surgery, NS Medical & Healthcare Service General Incorporation Foundation, Wakayama, Japan.
authors: Nobuo
authors: N
authors: Morita
authors: Oral and Maxillofacial Surgery II, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan.
authors: Yoshiaki
authors: Y
authors: Yura
authors: Departments of Molecular and Cellular Biochemistry, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan.
authors: Riko
authors: R
authors: Nishimura
authors: Departments of Molecular and Cellular Biochemistry, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan. toshyone@iu.edu.|Division of Hematology/Oncology, Department of Medicine, Indiana University School of Medicine, Walther Hall, R3-C321D, 980 W Walnut St, Indianapolis, IN, 46202, USA. toshyone@iu.edu.
authors: Toshiyuki
authors: T
authors: Yoneda
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D005353:Fibronectins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006258:Head and Neck Neoplasms / Q000473:pathology*; D064593:Heterografts; D006801:Humans; D007150:Immunohistochemistry; D042583:Lymphangiogenesis / Q000502:physiology; D008207:Lymphatic Metastasis / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D009062:Mouth Neoplasms / Q000473:pathology*; D060888:Real-Time Polymerase Chain Reaction; D000077195:Squamous Cell Carcinoma of Head and Neck; D042582:Vascular Endothelial Growth Factor C / Q000096:biosynthesis*
chemical_list: C067625:FN1 protein, human; D005353:Fibronectins; D042582:Vascular Endothelial Growth Factor C
keywords: Differential metastasis array; E-cadherin; Epithelial-mesenchymal transition; Focal adhesion kinase; Short hairpin RNA