Connective tissue growth factor/CCN2-null mouse embryonic fibroblasts retain intact transforming growth factor-{beta} responsiveness
Abstract
Background: The matricellular protein connective tissue growth factor (CCN2) has been implicated in pathological fibrosis, but its physiologic role remains elusive. In vitro, transforming growth factor-{beta} (TGF-{beta}) induces CCN2 expression in mesenchymal cells. Because CCN2 can enhance profibrotic responses elicited by TGF-{beta}, it has been proposed that CCN2 functions as an essential downstream signaling mediator for TGF-{beta}. To explore this notion, we characterized TGF-{beta}-induced activation of fibroblasts from CCN2-null (CCN2{sup -/-}) mouse embryos. Methods: The regulation of CCN2 expression was examined in vivo in a model of fibrosis induced by bleomycin. Cellular TGF-{beta} signal transduction and regulation of collagen gene expression were examined in CCN2{sup -/-} MEFs by immunohistochemistry, Northern, Western and RT-PCR analysis, immunocytochemistry and transient transfection assays. Results: Bleomycin-induced skin fibrosis in the mouse was associated with substantial CCN2 up-regulation in lesional fibroblasts. Whereas in vitro proliferation rate of CCN2{sup -/-} MEFs was markedly reduced compared to wild type MEFs, TGF-{beta}-induced activation of the Smad pathways, including Smad2 phosphorylation, Smad2/3 and Smad4 nuclear accumulation and Smad-dependent transcriptional responses, were unaffected by loss of CCN2. The stimulation of COL1A2 and fibronectin mRNA expression and promoter activity, and of corresponding protein levels, showed comparable time and dose-response in wild typemore »
- Authors:
-
- Division of Rheumatology, Northwestern University Feinberg School of Medicine, 240 E Huron Street, Chicago, IL 60611 (United States)
- OH/UCLA Department of Orthopedic Surgery, David Geffen School of Medicine, UCLA, Los Angeles, CA (United States)
- Publication Date:
- OSTI Identifier:
- 21045950
- Resource Type:
- Journal Article
- Journal Name:
- Experimental Cell Research
- Additional Journal Information:
- Journal Volume: 314; Journal Issue: 5; Other Information: DOI: 10.1016/j.yexcr.2007.12.010; PII: S0014-4827(07)00584-8; Copyright (c) 2007 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0014-4827
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 60 APPLIED LIFE SCIENCES; ACTIN; BLEOMYCIN; COLLAGEN; CONNECTIVE TISSUE; FIBROBLASTS; FIBROSIS; GROWTH FACTORS; IN VITRO; MICE; PHOSPHORYLATION; POLYMERASE CHAIN REACTION; SKIN; STIMULATION
Citation Formats
Mori, Yasuji, Hinchcliff, Monique, Wu, Minghua, Warner-Blankenship, Matthew, Lyons, Karen M, and Varga, John. Connective tissue growth factor/CCN2-null mouse embryonic fibroblasts retain intact transforming growth factor-{beta} responsiveness. United States: N. p., 2008.
Web. doi:10.1016/j.yexcr.2007.12.010.
Mori, Yasuji, Hinchcliff, Monique, Wu, Minghua, Warner-Blankenship, Matthew, Lyons, Karen M, & Varga, John. Connective tissue growth factor/CCN2-null mouse embryonic fibroblasts retain intact transforming growth factor-{beta} responsiveness. United States. https://doi.org/10.1016/j.yexcr.2007.12.010
Mori, Yasuji, Hinchcliff, Monique, Wu, Minghua, Warner-Blankenship, Matthew, Lyons, Karen M, and Varga, John. 2008.
"Connective tissue growth factor/CCN2-null mouse embryonic fibroblasts retain intact transforming growth factor-{beta} responsiveness". United States. https://doi.org/10.1016/j.yexcr.2007.12.010.
@article{osti_21045950,
title = {Connective tissue growth factor/CCN2-null mouse embryonic fibroblasts retain intact transforming growth factor-{beta} responsiveness},
author = {Mori, Yasuji and Hinchcliff, Monique and Wu, Minghua and Warner-Blankenship, Matthew and Lyons, Karen M and Varga, John},
abstractNote = {Background: The matricellular protein connective tissue growth factor (CCN2) has been implicated in pathological fibrosis, but its physiologic role remains elusive. In vitro, transforming growth factor-{beta} (TGF-{beta}) induces CCN2 expression in mesenchymal cells. Because CCN2 can enhance profibrotic responses elicited by TGF-{beta}, it has been proposed that CCN2 functions as an essential downstream signaling mediator for TGF-{beta}. To explore this notion, we characterized TGF-{beta}-induced activation of fibroblasts from CCN2-null (CCN2{sup -/-}) mouse embryos. Methods: The regulation of CCN2 expression was examined in vivo in a model of fibrosis induced by bleomycin. Cellular TGF-{beta} signal transduction and regulation of collagen gene expression were examined in CCN2{sup -/-} MEFs by immunohistochemistry, Northern, Western and RT-PCR analysis, immunocytochemistry and transient transfection assays. Results: Bleomycin-induced skin fibrosis in the mouse was associated with substantial CCN2 up-regulation in lesional fibroblasts. Whereas in vitro proliferation rate of CCN2{sup -/-} MEFs was markedly reduced compared to wild type MEFs, TGF-{beta}-induced activation of the Smad pathways, including Smad2 phosphorylation, Smad2/3 and Smad4 nuclear accumulation and Smad-dependent transcriptional responses, were unaffected by loss of CCN2. The stimulation of COL1A2 and fibronectin mRNA expression and promoter activity, and of corresponding protein levels, showed comparable time and dose-response in wild type and CCN2{sup -/-} MEFs, whereas stimulation of alpha smooth muscle actin and myofibroblast transdifferentiation showed subtle impairment in MEFs lacking CCN2. Conclusion: Whereas endogenous CCN2 plays a role in regulation of proliferation and TGF-{beta}-induced myofibroblast transdifferentiation, it appears to be dispensable for Smad-dependent stimulation of collagen and extracellular matrix synthesis in murine embryonic fibroblasts.},
doi = {10.1016/j.yexcr.2007.12.010},
url = {https://www.osti.gov/biblio/21045950},
journal = {Experimental Cell Research},
issn = {0014-4827},
number = 5,
volume = 314,
place = {United States},
year = {Mon Mar 10 00:00:00 EDT 2008},
month = {Mon Mar 10 00:00:00 EDT 2008}
}