Increased expression of matrilin‐3 not only in osteoarthritic articular cartilage but also in cartilage‐forming tumors, and down‐regulation of SOX9 via epidermal growth factor domain 1–dependent signaling

• Published in: Arthritis and rheumatism 2008-09, Vol.58 (9), p.2798-2808
• Main Authors: Vincourt, Jean‐Baptiste, Vignaud, Jean‐Michel, Lionneton, Frédéric, Sirveaux, François, Kawaki, Harumi, Marchal, Sophie, Lomazzi, Sandra, Plénat, François, Guillemin, François, Netter, Patrick, Takigawa, Masaharu, Mainard, Didier, Magdalou, Jacques
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Aged; Biological and medical sciences; Blotting, Western; Bone Neoplasms - genetics; Bone Neoplasms - metabolism; Cartilage, Articular - metabolism; Cell Differentiation - genetics; Cells, Cultured; Child; Child, Preschool; Chondrocytes - cytology; Chondrocytes - metabolism; Chondroma - genetics; Chondroma - metabolism; Chondrosarcoma - genetics; Chondrosarcoma - metabolism; Diseases of the osteoarticular system; Down-Regulation; Epidermal Growth Factor - genetics; Epidermal Growth Factor - metabolism; Extracellular Matrix Proteins - genetics; Extracellular Matrix Proteins - metabolism; Female; Flow Cytometry; Humans; Immunohistochemistry; Male; Matrilin Proteins; Medical sciences; Middle Aged; Miscellaneous. Osteoarticular involvement in other diseases; Osteoarthritis; Osteoarthritis - genetics; Osteoarthritis - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; Signal Transduction - genetics; SOX9 Transcription Factor - genetics; SOX9 Transcription Factor - metabolism
• ISSN: 0004-3591; 1529-0131
• DOI: 10.1002/art.23761

Objective To identify regulators of the cartilaginous phenotype, on the basis of their differential expression in human conventional chondrogenic tumors compared with articular cartilage. Methods Differential proteomics analysis revealed matrilin‐3 (MATN3) as a candidate regulator of the cartilaginous phenotype. Its capacity to modulate gene expression was investigated in human HCS‐2/8 chondrosarcoma cells and transfected chondrocytes, using cell culture fractionation, reverse transcription–polymerase chain reaction, and Western blot analyses. Results Increased expression of the cartilage‐specific matrix protein MATN3 was specifically observed in enchondromas and conventional chondrosarcomas. A substantial fraction of MATN3 was found in cytoplasmic structures of tumor cells, as demonstrated by immunohistochemistry. Analyses of intracellular MATN3 revealed that it corresponded to an imperfectly maturated MATN3 polypeptide, both in HCS‐2/8 human chondrosarcoma cells and in transfected human chondrocytes. Moderately increased expression of MATN3 resulted in its intracellular retention. Antibody‐mediated blockade of soluble, extracellular MATN3 in HCS‐2/8 cell cultures resulted in increased expression of MATN3 and the chondrogenic transcription factor SOX9. Conversely, increased ectopic expression of MATN3 resulted in decreased expression of MATN3 and SOX9 in primary chondrocytes, while a mutant MATN3 lacking its first epidermal growth factor (EGF)–like domain failed to down‐regulate SOX9. Conclusion Aberrant expression and processing of MATN3 are hallmarks of conventional cartilaginous neoplasms. A particular step in the maturation of MATN3 limits its processing through the secretion machinery, resulting in its intracellular accumulation upon increased expression. Soluble, secreted MATN3, however, down‐regulates SOX9 at the messenger RNA and protein levels. The first EGF‐like domain of MATN3 is a critical determinant of its regulatory activity toward SOX9. These activities of MATN3 suggest that its increased expression in osteoarthritis might contribute to the degeneration of articular cartilage.