PMID- 29149180 OWN - NLM STAT- MEDLINE DCOM- 20171226 LR - 20181113 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 12 IP - 11 DP - 2017 TI - Roles of VEGF-Flt-1 signaling in malignant behaviors of oral squamous cell carcinoma. PG - e0187092 LID - 10.1371/journal.pone.0187092 [doi] LID - e0187092 AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is a highly specific signaling protein for vascular endothelial cells that plays a critical role in tumor growth and invasion through angiogenesis, and may contribute to cell migration and activation of pre-osteoclasts, osteoclasts and some tumor cells. OBJECTIVES: We aimed to clarify the detailed roles of VEGF-Flt-1 signaling in bone invasion of oral squamous cell carcinoma (OSCC) cells. RESULTS: Forty-two (42) of 54 cases with gingival SCC (77.8%) strongly expressed VEGF, and had a significantly increased number of Flt-1+ osteoclasts (p<0.01) and more aggressive bone invasion (p<0.05). PlGF, a ligand of Flt-1, induced osteoclastogenesis in single culture of bone marrow cells (BMCs), and inhibition of Flt-1-signaling by VEGF tyrosine kinase inhibitor and It's down stream (Akt and ERK1/2) inhibitos reduced osteoclastogenesis in PlGF-stimulated BMCs (p<0.01). In molecular level, PlGF stimulation significantly upregulated RANKL expression in Flt-1-expressing HSC2 cells via phosphorylation of Akt and ERK1/2. In the co-culture of VEGF-producing HSC2 cells and BMCs, number of TRAP-positive osteoclasts markedly increased (p<0.01). The osteoclastogenesis was significantly inhibited by RANKL-neutralizing antibody (p<0.01) as well as by VEGF tyrosine kinase inhibitor (p<0.01) and it's downstream (Akt and ERK1/2) inhibitors (p<0.01, p<0.05, respectively). CONCLUSION: VEGF-Flt-1 signaling induces osteoclastogenesis in OSCC through two possible ways: 1) VEGF produced from OSCC cells can directly stimulate the Flt-1 pathway in preosteoclasts to induce migration to future bone resorbing area and differentiation into osteoclasts, and 2) VEGF-Flt-1 signaling upregulates RANKL expression in OSCC cells, which indirectly leads to osteoclast differentiation. Therefore, blocking of the VEGF-Flt-1 signaling may help inhibit bone invasion of OSCC. FAU - Subarnbhesaj, Ajiravudh AU - Subarnbhesaj A AD - Department of Oral and Maxillofacial Pathobiology, School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. FAU - Miyauchi, Mutsumi AU - Miyauchi M AUID- ORCID: 0000-0002-7428-9703 AD - Department of Oral and Maxillofacial Pathobiology, School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. FAU - Chanbora, Chea AU - Chanbora C AD - Department of Oral and Maxillofacial Pathobiology, School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. FAU - Mikuriya, Aki AU - Mikuriya A AD - Department of Oral and Maxillofacial Pathobiology, School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. FAU - Nguyen, Phuong Thao AU - Nguyen PT AD - Department of Global Dental Medicine and Pharmacy at Ho Chi Minh city, Integrated Health Sciences, School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. FAU - Furusho, Hisako AU - Furusho H AD - Department of Oral and Maxillofacial Pathobiology, School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. FAU - Ayuningtyas, Nurina Febriyanti AU - Ayuningtyas NF AD - Department of Oral and Maxillofacial Pathobiology, School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. FAU - Fujita, Minoru AU - Fujita M AD - Department of Oral and Maxillofacial Radiology, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. FAU - Toratani, Shigeaki AU - Toratani S AD - Department of Molecular Oral Medicine and Maxillofacial Surgery, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima University, Hiroshima, Japan. FAU - Takechi, Masaaki AU - Takechi M AD - Department of Oral and Maxillofacial Surgery, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. FAU - Niida, Shumpei AU - Niida S AD - Biobank, Medical Genome Center, National Center for Geriatrics and Gerontology, Obu, Japan. FAU - Takata, Takashi AU - Takata T AD - Department of Oral and Maxillofacial Pathobiology, School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan. LA - eng PT - Journal Article DEP - 20171117 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (Vascular Endothelial Growth Factor A) RN - EC 2.7.10.1 (Vascular Endothelial Growth Factor Receptor-1) SB - IM MH - Bone Neoplasms/secondary MH - Carcinoma, Squamous Cell/metabolism/*pathology MH - Cell Line, Tumor MH - Coculture Techniques MH - Gingiva/pathology MH - Humans MH - Mouth Neoplasms/metabolism/*pathology MH - Neoplasm Invasiveness MH - *Signal Transduction MH - Vascular Endothelial Growth Factor A/*metabolism MH - Vascular Endothelial Growth Factor Receptor-1/*metabolism PMC - PMC5693288 COIS- Competing Interests: The authors have declared that no competing interests exist. EDAT- 2017/11/18 06:00 MHDA- 2017/12/27 06:00 PMCR- 2017/11/17 CRDT- 2017/11/18 06:00 PHST- 2017/06/30 00:00 [received] PHST- 2017/10/15 00:00 [accepted] PHST- 2017/11/18 06:00 [entrez] PHST- 2017/11/18 06:00 [pubmed] PHST- 2017/12/27 06:00 [medline] PHST- 2017/11/17 00:00 [pmc-release] AID - PONE-D-17-19205 [pii] AID - 10.1371/journal.pone.0187092 [doi] PST - epublish SO - PLoS One. 2017 Nov 17;12(11):e0187092. doi: 10.1371/journal.pone.0187092. eCollection 2017.