TNF-α induces matrix metalloproteinase-9-dependent soluble intercellular adhesion molecule-1 release via TRAF2-mediated MAPKs and NF-κB activation in osteoblast-like MC3T3-E1 cellsReportar como inadecuado

TNF-α induces matrix metalloproteinase-9-dependent soluble intercellular adhesion molecule-1 release via TRAF2-mediated MAPKs and NF-κB activation in osteoblast-like MC3T3-E1 cells - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Journal of Biomedical Science

, 21:12

First Online: 05 February 2014Received: 14 September 2013Accepted: 28 January 2014DOI: 10.1186-1423-0127-21-12

Cite this article as: Tsai, CL., Chen, WC., Hsieh, HL. et al. J Biomed Sci 2014 21: 12. doi:10.1186-1423-0127-21-12


BackgroundMatrix metalloproteinase-9 MMP-9 has been shown to be induced by cytokines including TNF-α and may contribute to bone inflammatory diseases. However, the mechanisms underlying MMP-9 expression induced by TNF-α in MC3T3-E1 cells remain unclear.

ResultsWe applied gelatin zymography, Western blot, RT-PCR, real-time PCR, selective pharmacological inhibitors of transcription actinomycin D, Act.D, translation cycloheximide, CHI, c-Src PP1, MEK1-2 U0126, p38 MAPK SB202190, JNK1-2 SP600125, and NF-κB Bay11-7082, respective siRNAs transfection, promoter assay, immunofluorescence staining, and ELISA to investigate the MMP-9 expression and soluble ICAM-1 sICAM-1 release induced by TNF-α in MC3T3-E1 cells. Here we demonstrated that TNF-α-induced MMP-9 expression was attenuated by Act.D, CHI, PP1, U0126, SB202190, SP600125, and Bay11-7082, and by the transfection with siRNAs for ERK2, p38 MAPK, and JNK2. TNF-α-stimulated TNFR1, TRAF2, and c-Src complex formation was revealed by immunoprecipitation and Western blot. Furthermore, TNF-α-stimulated NF-κB phosphorylation and translocation were blocked by Bay11-7082, but not by PP1, U0126, SB202190, or SP600125. TNF-α time-dependently induced MMP-9 promoter activity which was also inhibited by PP1, U0126, SB202190, SP600125, or Bay11-7082. Up-regulation of MMP-9 was associated with the release of sICAM-1 into the cultured medium, which was attenuated by the pretreatment with MMP-2-9i, an MMP-9 inhibitor.

ConclusionsIn this study, we demonstrated that TNF-α up-regulates MMP-9 expression via c-Src, MAPKs, and NF-κB pathways. In addition, TNF-α-induced MMP-9 expression may contribute to the production of sICAM-1 by MC3T3-E1 cells. The interplay between MMP-9 expression and sICAM-1 release may exert an important role in the regulation of bone inflammatory diseases.

KeywordsTNF-α MMP-9 Soluble ICAM-1 MAPKs Osteoblast-like MC3T3-E1 cells AbbreviationsTNF-αTumor necrosis factor-α

MMP-9Matrix metalloproteinase-9

MC3T3 cellsMouse osteoblast-like cells

α-MEMMinimal essential medium-alpha

FBSFetal bovine serum

ECLEnhanced chemiluminescence

BCABicinchoninic acid

PBSPhosphate-buffered saline

BCABicinchoninic acid

TNFR1TNF receptor l

TRAF2TNF receptor-associated factor 2

MAPKsMitogen-activated protein kinases

ERKExtracellular-regulated protein kinase

JNKc-Jun-N-terminal kinase

sICAM-1Soluble intercellular adhesion molecule-1


siRNASmall interfering RNA

RT-PCRReverse transcription-polymerase chain reaction

NF-κBNuclear factor-kappa B.

Electronic supplementary materialThe online version of this article doi:10.1186-1423-0127-21-12 contains supplementary material, which is available to authorized users.

An erratum to this article is available at

Download fulltext PDF

Autor: Chia-Lan Tsai - Wei-Chung Chen - Hsi-Lung Hsieh - Pei-Ling Chi - Li-Der Hsiao - Chuen-Mao Yang


Documentos relacionados