Effects of Mutant Cartilage Oligomeric Matrix Protein on the Synthesis of Extracellular Matrix in the Swarm Rat Chondrosarcoma Cell Line.
- Author:
Hyun Woo KIM
1
;
Mun Soon PARK
;
Woo Suk SONG
;
Sun Young KONG
;
Kyoo Ho SHIN
;
Soo Bong HAHN
;
Hui Wan PARK
Author Information
1. Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, Korea. hwpark@yumc.yonsei.ac.kr
- Publication Type:Original Article
- Keywords:
Mutant COMP;
Extracellular matrix
- MeSH:
Animals;
Cartilage Oligomeric Matrix Protein*;
Cartilage*;
Cell Line*;
Chondrocytes;
Chondrosarcoma*;
Collagen;
Extracellular Matrix*;
Humans;
Proteoglycans;
Rats*;
Sepharose
- From:Journal of Korean Orthopaedic Research Society
2003;6(1):47-54
- CountryRepublic of Korea
- Language:Korean
-
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-II collagen detection among the cell line expressing mutant COMP and control cell lines. Histochemical staining of sulfated proteoglycans with safranin-O 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.