변이 연골 과소기질 단백질이 Swarm Rat 연골육종세포주의 세포외 기질 생성에 미치는 영향

Abstract

Purpose: To investigate the effects of mutated cartilage oligomeric matrix protein (COMP) on the synthesis of cartilage-specific major matrix proteins of Swarm rat chondrosarcoma chondrocytes. Materials and Methods: The Swarm rat chondrosarcoma chondrocytes transfected with chimeric construct consisting of a mutant gene of human COMP and an amino acid FLAG tag sequence were cultured in agarose gel. Formation of extracellular proteoglycan and type-Ⅱ collagen of the cells were evaluated by immunohistochemical staining and measuring 35S-sulfate incorporation. Results: No difference was observed in type-ll collagen detection among the cell line expressing mutant COMP and control cell lines. Histochemical staining of sulfated proteoglycans with safranin-0 showed lower amounts of proteoglycans were incorporated into the extracellular matrix of chondrocytes transfected with mutant gene. 35S-sulfate incorporation into the cell/matrix fractions demonstrates marked lower radiolabel incorporation compared to control cells. Conclusion: Mutation of COMP impacts the processing of proteoglycans rather than type-Ⅱ collagen in three-dimensional culture of Swarm rat chondrosarcoma chondrocytes.