{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,28]],"date-time":"2025-08-28T12:01:30Z","timestamp":1756382490457,"version":"3.40.5"},"reference-count":52,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2022,2,15]],"date-time":"2022-02-15T00:00:00Z","timestamp":1644883200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["614206"],"award-info":[{"award-number":["614206"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Prostate"],"published-print":{"date-parts":[[2022,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Metastatic prostate cancer lesions in the skeleton are frequently characterized by excessive formation of bone. Prostate cancer cells secrete factors, including serine proteases, that are capable of influencing the behavior of surrounding cells. Some of these proteases activate protease\u2010activated receptor\u20102 (PAR<jats:sub>2<\/jats:sub>), which is expressed by osteoblasts (bone\u2010forming cells) and precursors of osteoclasts (bone\u2010resorbing cells). The aim of the current study was to investigate a possible role for PAR<jats:sub>2<\/jats:sub> in regulating the behavior of bone cells exposed to metastatic prostate cancer cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The effect of medium conditioned by the PC3, DU145, and MDA\u2010PCa\u20102b prostate cancer cell lines was investigated in assays of bone cell function using cells isolated from wildtype and PAR<jats:sub>2<\/jats:sub>\u2010null mice. Osteoclast differentiation was assessed by counting tartrate\u2010resistant acid phosphatase\u2010positive multinucleate cells in bone marrow cultured in osteoclastogenic medium. Osteoblasts were isolated from calvariae of neonatal mice, and BrdU incorporation was used to assess their proliferation. Assays of alkaline phosphatase activity and quantitative PCR analysis of osteoblastic gene expression were used to assess osteoblast differentiation. Responses of osteoblasts to medium conditioned by MDA\u2010PCa\u20102b cells were analyzed by RNAseq.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Conditioned medium (CM) from all three cell lines inhibited osteoclast differentiation independently of PAR<jats:sub>2<\/jats:sub>. Media from PC3 and DU145 cells had no effect on assays of osteoblast function. Medium conditioned by MDA\u2010PCa\u20102b cells stimulated BrdU incorporation in both wildtype and PAR<jats:sub>2<\/jats:sub>\u2010null osteoblasts but increased alkaline phosphatase activity and <jats:italic>Runx2<\/jats:italic> and <jats:italic>Col1a1<\/jats:italic> expression in wildtype but not PAR<jats:sub>2<\/jats:sub>\u2010null cells. Functional enrichment analysis of RNAseq data identified enrichment of multiple gene ontology terms associated with lysosomal function in both wildtype and PAR<jats:sub>2<\/jats:sub>\u2010null cells in response to MDA\u2010PCa\u20102b\u2010CM. Analysis of individual genes identified osteogenesis\u2010associated genes that were either upregulated by MDA\u2010PCa\u20102b\u2010CM selectively in wildtype cells or downregulated selectively in PAR<jats:sub>2<\/jats:sub>\u2010null cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Factors secreted by prostate cancer cells influence bone cell behavior through both PAR<jats:sub>2<\/jats:sub>\u2010dependent and \u2010independent mechanisms. Both PAR<jats:sub>2<\/jats:sub>\u2010independent suppression of osteoclast differentiation and PAR<jats:sub>2<\/jats:sub>\u2010dependent stimulation of osteogenesis are likely to determine the nature of prostate cancer metastases in bone.<\/jats:p><\/jats:sec>","DOI":"10.1002\/pros.24316","type":"journal-article","created":{"date-parts":[[2022,2,15]],"date-time":"2022-02-15T14:19:44Z","timestamp":1644934784000},"page":"723-739","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Protease\u2010activated receptor\u20102 dependent and independent responses of bone cells to prostate cancer cell secretory products"],"prefix":"10.1002","volume":"82","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7844-2242","authenticated-orcid":false,"given":"Charles N.","family":"Pagel","sequence":"first","affiliation":[{"name":"Department of Veterinary Biosciences, Melbourne Veterinary School, Faculty of Veterinary and Agricultural 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