Comparative proteomic investigation of metastatic and non-metastatic osteosarcoma cells of human and canine origin

Abstract

Osteosarcoma is the most common bone cancer in dogs and people. In order to improve clinical outcomes, it is necessary to identify proteins that are differentially expressed by metastatic cells. Membrane bound proteins are responsible for multiple pro-metastatic functions. Therefore characterizing the differential expression of membranous proteins between metastatic and non-metastatic clonal variants will allow the discovery of druggable targets and consequently improve treatment methodology. The objective of this investigation was to systemically identify the membrane-associated proteomics of metastatic and non-metastatic variants of human and canine origin. Two clonal variants of divergent in vivo metastatic potential from human and canine origins were used. The plasma membranes were isolated and peptide fingerprinting was used to identify differentially expressed proteins. Selected proteins were further validated using western blotting, flow cytometry, confocal microscopy and immunohistochemistry. Over 500 proteins were identified for each cell line with nearly 40% of the proteins differentially regulated. Conserved between both species, metastatic variants demonstrated significant differences in expression of membrane proteins that are responsible for pro-metastatic functions. Additionally, CD147, CD44 and vimentin were validated using various biochemical techniques. Taken together, through a comparative proteomic approach we have identified several differentially expressed cell membrane proteins that will help in the development of future therapeutics.

Cluster of differentiation 44 promotes osteosarcoma progression in mice lacking the tumor suppressor Merlin

“…In the absence of tumor suppressors, CD44 repression might be released, prompting increased metastasis‐promoting functions. Numerous reports have implicated CD44 in the progression of osteosarcomas and other types of cancer 13,14,31,33‐35 . The action of CD44 in murine osteosarcoma appears to parallel osteosarcoma development in humans.…”

“…The action of CD44 in murine osteosarcoma appears to parallel osteosarcoma development in humans. CD44 is upregulated in metastatic subclones of human osteosarcoma and promotes lung metastasis formation in an allograft assay 33‐35 …”

Delineation of hypoxia-induced proteome shifts in osteosarcoma cells with different metastatic propensities

“…A recent glycoproteomic analysis performed on the isogenic human OS cell lines, HOS and 143B, also indicated upregulation of glycolysis/PPP proteins in the highly metastatic 143B cell line 55 . Noteworthy, a previous comparative proteomics study that focused on differentially expressed cell membrane proteins in metastatic and non-metastatic OS cells of human and canine origin showed that differentially expressed proteins associated with pro-metastatic functions were conserved between human and canine species 21 . Thus, targeting proteins involved in glycolysis in metastatic OS may inform new therapeutic strategies for canine as well as human osteosarcoma.…”

Protein Disulphide Isomerase A1 Is Involved in the Regulation of Breast Cancer Cell Adhesion and Transmigration via Lung Microvascular Endothelial Cells

“…The acquired MS/MS data were pre-processed with Mascot Distiller software (Matrix Science, London, UK), and a search was performed with the Mascot search engine against the human proteins. For the detection and semi quantitation of PDI isoforms and integrin isoforms, the exponentially modified protein abundance index (emPAI) calculation was performed [ 64 , 65 ]. The detailed procedures for proteomic studies are described in the Supplementary Materials and Methods .…”