Osteopontin expression in co-cultures of human squamous cell carcinoma-derived cells and osteoblastic cells and its effects on the neoplastic cell phenotype and osteoclastic activation.
This study evaluated the temporal expression of osteopontin (OPN) in co-cultures of human osteoblastic cells (SAOS-2) and oral squamous cell carcinoma (OSCC)-derived cells (SCC9) and examined the effects of osteoblast-derived OPN on the neoplastic cell phenotype. Additionally, the effects of these co-cultures on subsequent osteoclastic activity were explored. SCC9 cells were plated on Transwell® membranes that were either coated or not coated with Matrigel and were then co-cultured with SAOS-2 cells during the peak of OPN expression. SCC9 cells exposed to OPN-silenced SAOS-2 cultures and SCC9 cells cultured alone served as controls. SCC9 cells were quantitatively evaluated for cell adhesion, proliferation, migration, and invasion into Matrigel. The impact of co-culturing SAOS-2 and SCC9 cells on the resorptive capacity of U-937-derived osteoclastic cells was also investigated. Furthermore, a reciprocal induction of SAOS-2 and SCC9 cells in terms of OPN expression over the co-culture interval was identified. SAOS-2-secreted OPN altered the SCC9 cell phenotype, leading to enhanced cell adhesion and proliferation and higher Matrigel invasion. This invasion was also enhanced, albeit to a lesser degree, by co-culture with OPN-silenced SAOS-2 cells. Cell migration was not affected. Co-culture with SAOS-2 cells-mainly during the period of peak OPN expression-promoted over-expression of IL-6 and IL-8 by SCC9 cells and enhanced the resorptive capacity of osteoclastic cells. Taken together, these results suggest that osteoblast-derived OPN affects the interactions among OSCC-derived epithelial cells, osteoblasts, and osteoclasts, which could contribute to the process of bone destruction during bone invasion by OSCC.
Cell Culture Laboratory, Department of Morphology, Physiology and Basic Pathology, School of Dentistry of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, Monte Alegre, Ribeirão Preto, São Paulo, 14040-904, Brazil. novaesrp@yahoo.com.br.|Department of Oral Diagnosis, School of Dentistry of Piracicaba, State University of Campinas, Av. Limeira, 901, Vila Rezende, Piracicaba, São Paulo, 13414-903, Brazil. novaesrp@yahoo.com.br.
Lucas Novaes
LN
Teixeira
Cell Culture Laboratory, Department of Morphology, Physiology and Basic Pathology, School of Dentistry of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, Monte Alegre, Ribeirão Preto, São Paulo, 14040-904, Brazil.
Larissa Moreira Spinola
LM
de Castro Raucci
Cell Culture Laboratory, Department of Morphology, Physiology and Basic Pathology, School of Dentistry of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, Monte Alegre, Ribeirão Preto, São Paulo, 14040-904, Brazil.
Gabriela Caroline
GC
Alonso
Department of Oral Diagnosis, School of Dentistry of Piracicaba, State University of Campinas, Av. Limeira, 901, Vila Rezende, Piracicaba, São Paulo, 13414-903, Brazil.
Ricardo Della
RD
Coletta
Cell Culture Laboratory, Department of Morphology, Physiology and Basic Pathology, School of Dentistry of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, Monte Alegre, Ribeirão Preto, São Paulo, 14040-904, Brazil.
Adalberto Luiz
AL
Rosa
Cell Culture Laboratory, Department of Morphology, Physiology and Basic Pathology, School of Dentistry of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, Monte Alegre, Ribeirão Preto, São Paulo, 14040-904, Brazil. tambasco@usp.br.
Paulo Tambasco
PT
de Oliveira
D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002448:Cell Adhesion / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D018920:Coculture Techniques; D003094:Collagen; D016207:Cytokines / Q000378:metabolism; D004338:Drug Combinations; D004797:Enzyme-Linked Immunosorbent Assay; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007797:Laminin; D008856:Microscopy, Fluorescence; D010006:Osteoblasts / Q000378:metabolism* / Q000473:pathology; D010010:Osteoclasts / Q000378:metabolism* / Q000473:pathology; D053495:Osteopontin / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D011509:Proteoglycans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020298:U937 Cells
D016207:Cytokines; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D053495:Osteopontin; C059485:matrigel; D003094:Collagen
Bone invasion; Co-culture; Oral squamous cell carcinoma; Osteoblasts; Osteopontin