Abstract
The synthesis, processing, and secretion of human connective tissue growth factor (CTGF/Hcs24) in a human chondrocytic cell line, HCS-2/8, were analyzed immunochemically. By metabolic pulse-labeling, chasing, and subsequent immunoprecipitation analyses, active synthesis of CTGF was observed not only in growing HCS-2/8 cells, but also in confluent cells. However, secretion and processing of CTGF were found to be regulated differentially, depending upon the growth status. During phases of growth, HCS-2/8 cells released CTGF molecules immediately without sequestering them within the cell layer. In contrast, after the cells reached confluence, the secretion slowed, resulting in an accumulation of CTGF in the cells or extracellular matrices (ECMs). Also, in confluent cell layers, a 10 kDa protein that was reactive to an anti-CTGF serum was observed. This CTGF-related small protein was not detected immediately after labeling, but gradually appeared within 6 h after chase, which suggests its entity as a processed subfragment of CTGF. Surprisingly, the 10 kDa protein was stable even 48 h after synthesis, and was not released by ECM digestion, suggesting an intracellular maintenance and function. Taken together, the behavior of CTGF in HCS-2/8 cells is remarkably different from that reported in fibroblasts, which may represent unique roles for CTGF in the growth and differentiation of chondrocytes.
| Original language | English |
|---|---|
| Pages (from-to) | 155-161 |
| Number of pages | 7 |
| Journal | Bone |
| Volume | 29 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2001 |
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Keywords
- Cartilage
- Chondrocyte
- Connective tissue growth factor (CTGF)
- Extracellular matrix (ECM)
- HCS-2/8
ASJC Scopus subject areas
- Physiology
- Hematology
Cite this
Novel mode of processing and secretion of connective tissue growth factor/ecogenin (CTGF/Hcs24) in chondrocytic HCS-2/8 cells. / Kubota, Satoshi; Eguchi, Takanori; Shimo, T.; Nishida, Takashi; Hattori, Takako; Kondo, S.; Nakanishi, T.; Takigawa, Masaharu.
In: Bone, Vol. 29, No. 2, 2001, p. 155-161.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Novel mode of processing and secretion of connective tissue growth factor/ecogenin (CTGF/Hcs24) in chondrocytic HCS-2/8 cells
AU - Kubota, Satoshi
AU - Eguchi, Takanori
AU - Shimo, T.
AU - Nishida, Takashi
AU - Hattori, Takako
AU - Kondo, S.
AU - Nakanishi, T.
AU - Takigawa, Masaharu
PY - 2001
Y1 - 2001
N2 - The synthesis, processing, and secretion of human connective tissue growth factor (CTGF/Hcs24) in a human chondrocytic cell line, HCS-2/8, were analyzed immunochemically. By metabolic pulse-labeling, chasing, and subsequent immunoprecipitation analyses, active synthesis of CTGF was observed not only in growing HCS-2/8 cells, but also in confluent cells. However, secretion and processing of CTGF were found to be regulated differentially, depending upon the growth status. During phases of growth, HCS-2/8 cells released CTGF molecules immediately without sequestering them within the cell layer. In contrast, after the cells reached confluence, the secretion slowed, resulting in an accumulation of CTGF in the cells or extracellular matrices (ECMs). Also, in confluent cell layers, a 10 kDa protein that was reactive to an anti-CTGF serum was observed. This CTGF-related small protein was not detected immediately after labeling, but gradually appeared within 6 h after chase, which suggests its entity as a processed subfragment of CTGF. Surprisingly, the 10 kDa protein was stable even 48 h after synthesis, and was not released by ECM digestion, suggesting an intracellular maintenance and function. Taken together, the behavior of CTGF in HCS-2/8 cells is remarkably different from that reported in fibroblasts, which may represent unique roles for CTGF in the growth and differentiation of chondrocytes.
AB - The synthesis, processing, and secretion of human connective tissue growth factor (CTGF/Hcs24) in a human chondrocytic cell line, HCS-2/8, were analyzed immunochemically. By metabolic pulse-labeling, chasing, and subsequent immunoprecipitation analyses, active synthesis of CTGF was observed not only in growing HCS-2/8 cells, but also in confluent cells. However, secretion and processing of CTGF were found to be regulated differentially, depending upon the growth status. During phases of growth, HCS-2/8 cells released CTGF molecules immediately without sequestering them within the cell layer. In contrast, after the cells reached confluence, the secretion slowed, resulting in an accumulation of CTGF in the cells or extracellular matrices (ECMs). Also, in confluent cell layers, a 10 kDa protein that was reactive to an anti-CTGF serum was observed. This CTGF-related small protein was not detected immediately after labeling, but gradually appeared within 6 h after chase, which suggests its entity as a processed subfragment of CTGF. Surprisingly, the 10 kDa protein was stable even 48 h after synthesis, and was not released by ECM digestion, suggesting an intracellular maintenance and function. Taken together, the behavior of CTGF in HCS-2/8 cells is remarkably different from that reported in fibroblasts, which may represent unique roles for CTGF in the growth and differentiation of chondrocytes.
KW - Cartilage
KW - Chondrocyte
KW - Connective tissue growth factor (CTGF)
KW - Extracellular matrix (ECM)
KW - HCS-2/8
UR - http://www.scopus.com/inward/record.url?scp=0034897156&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034897156&partnerID=8YFLogxK
U2 - 10.1016/S8756-3282(01)00492-6
DO - 10.1016/S8756-3282(01)00492-6
M3 - Article
C2 - 11502477
AN - SCOPUS:0034897156
VL - 29
SP - 155
EP - 161
JO - Bone
JF - Bone
SN - 8756-3282
IS - 2
ER -