C-Src-dependent transactivation of PDGFR contributes to TNF-α-induced MMP-9 expression and functional impairment in osteoblasts

Chia Lan Tsai, Wei Chung Chen, I-Ta Lee, Pei Ling Chi, Shin Ei Cheng, Chuen Mao Yang

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Matrix metalloproteinases (MMPs), MMP-9 especially, have been shown to be induced by cytokines, including tumor necrosis factor-α (TNF-α) and may contribute to bone inflammatory diseases and postnatal bone modeling and remodeling. However, the mechanisms underlying MMP-9 expression induced by TNF-α in osteoblasts remain unclear. Here, we showed that in MC3T3-E1 cells, TNF-α induced MMP-9 gene expression determined by real-time PCR, zymography, and promoter assay. TNF-α-mediated responses were attenuated by pretreatment with the inhibitor of protein tyrosine kinase (PTK; genistein), c-Src (PP1), PDGFR (AG1296), PI3K (LY294002), Akt (SH-5), MEK1/2 (U0126), p38 MAPK (SB202190), JNK1/2 (SP600125), or AP-1 (Tanshinone IIA) and transfection with siRNA of c-Src, PDGFR, p85, Akt, c-Jun, or ATF2. Moreover, TNF-α also time-dependently stimulated phosphorylation of c-Src and PDGFR and c-Src/PDGFR complex formation, which were reduced by pretreatment with PP1 or AG1296. TNF-α-stimulated Akt phosphorylation was inhibited by genistein, PP1, AG1296, LY294002, or SH5. We further demonstrated that TNF-α stimulated ERK1/2, p38 MAPK, and JNK1/2 phosphorylation via a c-Src-dependent PDGFR/PI3K/Akt pathway. TNF-α stimulated AP-1 activation, including c-Jun and ATF2 phosphorylation and AP-1 transcription activity via MAPK-dependent pathways. In addition, TNF-α-induced MMP-9 promoter activity was mediated through an AP-1 binding domain of the MMP-9 promoter region. Finally, we found that up-regulation of MMP-9 contributes to MMP-mediated type I collagen degradation and osteoblasts detachment. These results suggested that TNF-α-induced MMP-9 expression is mediated through a c-Src-dependent PDGFR transactivation and PI3K/Akt cascade linking to MAPK-mediated activation of AP-1 (c-Jun/ATF2) and leading to functional impairment in osteoblasts.

Original languageEnglish
Pages (from-to)186-197
Number of pages12
JournalBone
Volume60
DOIs
Publication statusPublished - Mar 1 2014
Externally publishedYes

Fingerprint

Matrix Metalloproteinase 9
Osteoblasts
Transcriptional Activation
Tumor Necrosis Factor-alpha
Transcription Factor AP-1
Phosphatidylinositol 3-Kinases
Phosphorylation
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Genistein
p38 Mitogen-Activated Protein Kinases
Matrix Metalloproteinases
Bone Remodeling
Bone Diseases
Collagen Type I
Genetic Promoter Regions
Protein-Tyrosine Kinases
Small Interfering RNA
Transfection
Real-Time Polymerase Chain Reaction
Up-Regulation

Keywords

  • Cell-mediated type I collagen degradation
  • Cytokine
  • MMP-9
  • Osteoblast-like MC3T3-E1 cells
  • PDGFR transactivation

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Histology

Cite this

C-Src-dependent transactivation of PDGFR contributes to TNF-α-induced MMP-9 expression and functional impairment in osteoblasts. / Tsai, Chia Lan; Chen, Wei Chung; Lee, I-Ta; Chi, Pei Ling; Cheng, Shin Ei; Yang, Chuen Mao.

In: Bone, Vol. 60, 01.03.2014, p. 186-197.

Research output: Contribution to journalArticle

Tsai, Chia Lan ; Chen, Wei Chung ; Lee, I-Ta ; Chi, Pei Ling ; Cheng, Shin Ei ; Yang, Chuen Mao. / C-Src-dependent transactivation of PDGFR contributes to TNF-α-induced MMP-9 expression and functional impairment in osteoblasts. In: Bone. 2014 ; Vol. 60. pp. 186-197.
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abstract = "Matrix metalloproteinases (MMPs), MMP-9 especially, have been shown to be induced by cytokines, including tumor necrosis factor-α (TNF-α) and may contribute to bone inflammatory diseases and postnatal bone modeling and remodeling. However, the mechanisms underlying MMP-9 expression induced by TNF-α in osteoblasts remain unclear. Here, we showed that in MC3T3-E1 cells, TNF-α induced MMP-9 gene expression determined by real-time PCR, zymography, and promoter assay. TNF-α-mediated responses were attenuated by pretreatment with the inhibitor of protein tyrosine kinase (PTK; genistein), c-Src (PP1), PDGFR (AG1296), PI3K (LY294002), Akt (SH-5), MEK1/2 (U0126), p38 MAPK (SB202190), JNK1/2 (SP600125), or AP-1 (Tanshinone IIA) and transfection with siRNA of c-Src, PDGFR, p85, Akt, c-Jun, or ATF2. Moreover, TNF-α also time-dependently stimulated phosphorylation of c-Src and PDGFR and c-Src/PDGFR complex formation, which were reduced by pretreatment with PP1 or AG1296. TNF-α-stimulated Akt phosphorylation was inhibited by genistein, PP1, AG1296, LY294002, or SH5. We further demonstrated that TNF-α stimulated ERK1/2, p38 MAPK, and JNK1/2 phosphorylation via a c-Src-dependent PDGFR/PI3K/Akt pathway. TNF-α stimulated AP-1 activation, including c-Jun and ATF2 phosphorylation and AP-1 transcription activity via MAPK-dependent pathways. In addition, TNF-α-induced MMP-9 promoter activity was mediated through an AP-1 binding domain of the MMP-9 promoter region. Finally, we found that up-regulation of MMP-9 contributes to MMP-mediated type I collagen degradation and osteoblasts detachment. These results suggested that TNF-α-induced MMP-9 expression is mediated through a c-Src-dependent PDGFR transactivation and PI3K/Akt cascade linking to MAPK-mediated activation of AP-1 (c-Jun/ATF2) and leading to functional impairment in osteoblasts.",
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AU - Chi, Pei Ling

AU - Cheng, Shin Ei

AU - Yang, Chuen Mao

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N2 - Matrix metalloproteinases (MMPs), MMP-9 especially, have been shown to be induced by cytokines, including tumor necrosis factor-α (TNF-α) and may contribute to bone inflammatory diseases and postnatal bone modeling and remodeling. However, the mechanisms underlying MMP-9 expression induced by TNF-α in osteoblasts remain unclear. Here, we showed that in MC3T3-E1 cells, TNF-α induced MMP-9 gene expression determined by real-time PCR, zymography, and promoter assay. TNF-α-mediated responses were attenuated by pretreatment with the inhibitor of protein tyrosine kinase (PTK; genistein), c-Src (PP1), PDGFR (AG1296), PI3K (LY294002), Akt (SH-5), MEK1/2 (U0126), p38 MAPK (SB202190), JNK1/2 (SP600125), or AP-1 (Tanshinone IIA) and transfection with siRNA of c-Src, PDGFR, p85, Akt, c-Jun, or ATF2. Moreover, TNF-α also time-dependently stimulated phosphorylation of c-Src and PDGFR and c-Src/PDGFR complex formation, which were reduced by pretreatment with PP1 or AG1296. TNF-α-stimulated Akt phosphorylation was inhibited by genistein, PP1, AG1296, LY294002, or SH5. We further demonstrated that TNF-α stimulated ERK1/2, p38 MAPK, and JNK1/2 phosphorylation via a c-Src-dependent PDGFR/PI3K/Akt pathway. TNF-α stimulated AP-1 activation, including c-Jun and ATF2 phosphorylation and AP-1 transcription activity via MAPK-dependent pathways. In addition, TNF-α-induced MMP-9 promoter activity was mediated through an AP-1 binding domain of the MMP-9 promoter region. Finally, we found that up-regulation of MMP-9 contributes to MMP-mediated type I collagen degradation and osteoblasts detachment. These results suggested that TNF-α-induced MMP-9 expression is mediated through a c-Src-dependent PDGFR transactivation and PI3K/Akt cascade linking to MAPK-mediated activation of AP-1 (c-Jun/ATF2) and leading to functional impairment in osteoblasts.

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