Identification of STAT-1 as a molecular target of IGFBP-3 in the process of chondrogenesis

Anna Spagnoli, Monica Torello, Srivinasa R. Nagalla, William A. Horton, Patrick Pattee, Vivian Hwa, Francesco Chiarelli, Charles Roberts, Ronald (Ron) Rosenfeld

Research output: Contribution to journalArticle

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Abstract

The chondrogenesis process requires the ordered proliferation and differentiation of chondrocytes. Insulin-like growth factor-binding protein (IGFBP)-3, well characterized as the carrier of insulin-like growth factor (IGF), has been reported to have intrinsic bioactivity that is independent of IGF binding. The mechanisms involved in this IGF-independent action are still unclear. Using the RCJ3.1C5.18 chondrogenic cells, which in culture progresses from undifferentiated to terminally differentiated chondrocytes, we have shown previously that IGFBP-3 has an IGF-independent, antiproliferative effect in undifferentiated and early differentiated but not in terminally differentiated chondrocytes. In the present study, cDNA microarray analysis was used to screen for genes: 1) that were regulated by IGFBP-3 in early but not in terminally differentiated chondrocytes; 2) that were regulated specifically by IGFBP-3, but not by IGF-I; and 3) whose regulation was abolished by coincubation of IG-FBP-3 with IGF-I. Signal transducer and activator of transcription (STAT)-I was the gene that, fulfilling the screening criteria, exhibited the greatest up-regulation by IGFBP-3 (>40-fold). STAT-1 gene up-regulation was confirmed by Northern analysis of cells treated with IGFBP-3 or transfected with an IGFBP-3 expression vector. Remarkably, similar results were obtained when cells were transfected with an IGFBP-3 mutant unable to bind IGFs, definitively demonstrating the IGF-independent action of IGFBP-3. Consistent with the up-regulation of STAT-1 mRNA, IGFBP-3 also increased STAT-1 protein expression. Furthermore, both IGFBP-3 and the IGFBP-3 mutant induced STAT-1 phosphorylation and its nuclear localization. An antisense STAT-1 oligonucleotide abolished the IGF-independent cell apoptosis induced by IGFBP-3. We have demonstrated that STAT-1 is a major intracellular signaling and transcriptional target of the IGF-independent apoptotic effect of IGFBP-3 in chondrogenesis.

Original languageEnglish (US)
Pages (from-to)18860-18867
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number21
DOIs
StatePublished - May 24 2002
Externally publishedYes

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STAT1 Transcription Factor
Chondrogenesis
Insulin-Like Growth Factor Binding Protein 3
Somatomedins
Chondrocytes
Up-Regulation
Genes
Insulin-Like Growth Factor I
Replication Protein C
STAT Transcription Factors
Phosphorylation
Transcription
Microarray Analysis
Microarrays
Bioactivity
Oligonucleotide Array Sequence Analysis
Transducers

ASJC Scopus subject areas

  • Biochemistry

Cite this

Spagnoli, A., Torello, M., Nagalla, S. R., Horton, W. A., Pattee, P., Hwa, V., ... Rosenfeld, R. R. (2002). Identification of STAT-1 as a molecular target of IGFBP-3 in the process of chondrogenesis. Journal of Biological Chemistry, 277(21), 18860-18867. https://doi.org/10.1074/jbc.M200218200

Identification of STAT-1 as a molecular target of IGFBP-3 in the process of chondrogenesis. / Spagnoli, Anna; Torello, Monica; Nagalla, Srivinasa R.; Horton, William A.; Pattee, Patrick; Hwa, Vivian; Chiarelli, Francesco; Roberts, Charles; Rosenfeld, Ronald (Ron).

In: Journal of Biological Chemistry, Vol. 277, No. 21, 24.05.2002, p. 18860-18867.

Research output: Contribution to journalArticle

Spagnoli, A, Torello, M, Nagalla, SR, Horton, WA, Pattee, P, Hwa, V, Chiarelli, F, Roberts, C & Rosenfeld, RR 2002, 'Identification of STAT-1 as a molecular target of IGFBP-3 in the process of chondrogenesis', Journal of Biological Chemistry, vol. 277, no. 21, pp. 18860-18867. https://doi.org/10.1074/jbc.M200218200
Spagnoli A, Torello M, Nagalla SR, Horton WA, Pattee P, Hwa V et al. Identification of STAT-1 as a molecular target of IGFBP-3 in the process of chondrogenesis. Journal of Biological Chemistry. 2002 May 24;277(21):18860-18867. https://doi.org/10.1074/jbc.M200218200
Spagnoli, Anna ; Torello, Monica ; Nagalla, Srivinasa R. ; Horton, William A. ; Pattee, Patrick ; Hwa, Vivian ; Chiarelli, Francesco ; Roberts, Charles ; Rosenfeld, Ronald (Ron). / Identification of STAT-1 as a molecular target of IGFBP-3 in the process of chondrogenesis. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 21. pp. 18860-18867.
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AU - Pattee, Patrick

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