Characterization of bone resorption in novel in vitro and in vivo models of oral squamous cell carcinoma

Martin, Chelsea K.; Dirksen, Wessel P.; Shu, Sherry T.; Werbeck, Jillian L.; Thudi, Nanda K.; Yamaguchi, Mamoru; Wolfe, Tobie D.; Heller, Kristin N.; Rosol, Thomas J.

doi:10.1016/j.oraloncology.2011.12.012

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Characterization of bone resorption in novel in vitro and in vivo models of oral squamous cell carcinoma

Chelsea K. Martin
- Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, USA
Wessel P. Dirksen
- Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, USA
Sherry T. Shu
- Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, PA, USA
Jillian L. Werbeck
- Cancer Biology and Genetics, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
Nanda K. Thudi
- Department of Radiation Oncology, University of Alabama, Birmingham, AL, USA
Mamoru Yamaguchi
- Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, USA
Tobie D. Wolfe
- Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, USA
Kristin N. Heller
- Department of Molecular, Cellular and Developmental Biology, The Ohio State University, Columbus, OH, USA
Thomas J. Rosol
- Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, USA
- Corresponding author. Tel.: +1 614 292 4265.

Received 22 August 2011; received in revised form 12 December 2011; accepted 23 December 2011. published online 24 January 2012.
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Summary

Oral squamous cell carcinoma (OSCC) is the most commonly diagnosed oral malignancy in humans and cats and frequently invades bone. The objective of this study was to determine if feline OSCC serves as a relevant model of human OSCC in terms of osteolytic behavior and expression of bone resorption agonists. Novel feline OSCC cell lines (SCCF2 and SCCF3) were derived from spontaneous carcinomas. Gene expression and osteolytic behavior were compared to an established feline OSCC cell line (SCCF1) and three human OSCC cell lines (UMSCC-12, A253 and SCC25). Interaction of OSCC with bone and murine pre-osteoblasts (MC3T3) was investigated using in vitro co-culture techniques. In vivo bioluminescent imaging, Faxitron radiography and microscopy were used to measure xenograft growth and bone invasion in nude mice. Human and feline OSCC expressing the highest levels of parathyroid hormone-related protein (PTHrP) were associated with in vitro and in vivo bone resorption and osteoclastogenesis. MC3T3 cells had increased receptor activator of nuclear factor κB ligand (RANKL) expression and reduced osteoprotegerin (OPG) expression in conditioned medium from bone-invasive SCCF2 cells compared to minimally bone invasive SCCF3 cells, which was partially reversed with a neutralizing anti-PTHrP antibody. Human and feline OSCC cells cultured in bone-conditioned medium had increased PTHrP secretion and proliferation. Feline OSCC-induced bone resorption was associated with tumor cell secretion of PTHrP and with increased RANKL:OPG expression ratio in mouse preosteoblasts. Bone-CM increased OSCC proliferation and secretion of PTHrP. The preclinical models of feline OSCC recapitulated the bone-invasive phenotype characteristic of spontaneous OSCC and will be useful to future preclinical and mechanistic studies of bone invasive behavior.

Keywords: Oral squamous cell carcinoma, Osteoclast, Bone invasion, Parathyroid hormone-related protein, Mouse model, Feline