ArticleGene2006
Identification of cis and trans-acting transcriptional regulators in chondroinduced fibroblasts from the pre-phenotypic gene expression profile.
Article in Gene, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 13 citations in OpenAlex.
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- Recent progress in histochemistry and cell biology.Histochemistry and cell biology · 2012Review
- A proteomic approach for identification and localization of the pericellular components of chondrocytes.Histochemistry and cell biology · 2011Article
- Arterial injury promotes medial chondrogenesis in Sm22 knockout mice.Cardiovascular research · 2011Article
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- A cis-regulatory site downregulates PTHLH in translocation t(8;12)(q13;p11.2) and leads to Brachydactyly Type E.Human molecular genetics · 2010Article
- Regulation of the human SOX9 promoter by Sp1 and CREB.Experimental cell research · 2007Article
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1 author at 1 institution in 1 country.
Funding
Abstract
Cell differentiation is regulated via expression of successive sets of genes. In an in vitro model of chondrocyte differentiation, human dermal fibroblasts (hDFs) cultured in collagen sponges are induced to express cartilage matrix genes after 7 days' culture with demineralized bone powder (DBP). A shift in expression of many other genes occurs within 3 days, before chondroblast phenotypic genes are detectable. In this study, the pre-chondrogenic gene expression profile was used as a starting point to derive information on transcriptional regulation of chondrocyte differentiation induced by DBP. Putative cis regulatory elements were identified by comparing promoter regions from three genes that are highly upregulated in chondroinduced hDFs (BIGH3, COL1A2, and FN1) [Zhou, S., Glowacki, J., Yates, K.E, 2004. Comparison of TGF-beta/BMP pathways signaled by demineralized bone powder and BMP-2 in human dermal fibroblasts. J. Bone Min. Res. 19, 1732-1741] and whose products are known to interact in the matrix [Kim, J.E., et al., 2002. Molecular properties of wild-type and mutant betaIG-H3 proteins. Investig. Ophthalmol. Vis. Sci. 43, 656-661]. The effect of DBP on nuclear protein binding to cis elements was measured with an array-based assay. Nuclear extracts from hDFs cultured in DBP/collagen sponges for 3 days showed increased binding to several cis elements belonging to the families that were identified by promoter analysis. Of note, those elements represented targets of both signal-activated and developmentally regulated transcription factors. Direct measurement of mRNAs showed increased gene expression of both types of transcription factors in chondroinduced hDFs, including NFKB2 (290% of control), RELA (160%), and GATA2 (190%). Moreover, DBP increased gene expression of chondrogenic transcription factors SOX9 (160% of control) and RUNX2 (180%). Immunoblot analysis showed that DBP increased both expression (200% of control) and phosphorylation (300%) of the Creb protein, a transcription factor that is downstream of several signal transduction pathways. Inhibition of protein kinase A, protein kinase C, or MAP kinase in hDFs cultured in DBP/collagen sponges reduced induction of BIGH3 to approximately 50% of control. These results suggest that both signal-activated and developmentally controlled transcriptional mechanisms contribute to chondroinduction of hDFs by DBP.
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