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J. Biol. Chem., Vol. 276, Issue 8, 5533-5540, February 23, 2001
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From the Chondrogenesis results from a complex equilibrium
between chondrocyte proliferation and differentiation. Insulin-like
growth factors (IGFs) have a crucial role in chondrogenesis, but their mechanisms of action are not well defined. IGF-binding protein-3 (IGFBP-3) is the major carrier for circulating IGFs in postnatal life,
and has been shown to have IGF-independent effects on proliferation of
several cancer cell lines. In this study, we have evaluated the
IGF-independent and -dependent effects of IGFBP-3 on
chondrocyte proliferation and the relationship of these effects with
chondrocyte differentiation stage. We used the RCJ3.1C5.18
nontransformed mesenchymal chondrogenic cell line, which, over 2 weeks
of culture, progresses through the differentiation pathway exhibited by
chondrocytes in the growth plate. We demonstrated that IGFBP-3
inhibited, in a dose-dependent manner (1-30
nM), the proliferation of chondroprogenitors and
early differentiated chondrocytes, stimulated by des-(1-3)-IGF-I and
longR3-IGF-I (IGF-I analogs with reduced affinity for
IGFBP-3), and by insulin and IGF-I. In terminally differentiated
chondrocytes, IGFBP-3 retained the ability to inhibit cell
proliferation stimulated by IGF-I, but had no effect on cell growth
stimulated by insulin, or des-(1-3)-IGF-I or longR3IGF-I.
By monolayer affinity cross-linking, we demonstrated a specific
IGFBP-3-associated cell-membrane protein of ~20 kDa. We determined
that IGFBP-3 has an antiproliferative effect on chondrocytes and, that
this effect is related to the differentiation process. In
chondroprogenitors and early differentiated chondrocytes, antiproliferative effect of IGFBP-3 is mainly IGF-independent, whereas,
following terminal differentiation this effect is
IGF-dependent.
Antiproliferative Effects of Insulin-like Growth Factor-binding
Protein-3 in Mesenchymal Chondrogenic Cell Line RCJ3.1C5.18
RELATIONSHIP TO DIFFERENTIATION STAGE*
§,,,
,, and
Department of Pediatrics, Oregon Health
Sciences University, Portland, Oregon 97201, the ¶ Research
Department, Shriners Hospital for Children, Portland, Oregon 97201, and
the Department of Pediatrics, University of Chieti,
66013 Chieti, Italy
*
This work was supported in part by a grant from the Medical
Research Foundation of Oregon (to A. S.), by a grant from
Pharmacia & Upjohn (to A. S.), and by National Institutes of
Health Grant CA58110 (to R. G. R.). This work was presented
in part at the 81st Annual Meeting of the Endocrine Society, June
12-15, 1999, San Diego, CA.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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