Regulation of mechanical stress-induced MMP-13 and ADAMTS-5 expression by RUNX-2 transcriptional factor in SW1353 chondrocyte-like cells

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Abstract

Objective: To investigate the mechanism of mechanical stress-induced expression and regulation of aggrecanases and examine the role of runt-related transcription factor 2 (RUNX-2) in chondrocyte-like cells. Methods: SW1353 cells were seeded onto stretch chambers at a concentration of 5×104cells/chamber, and a uni-axial cyclic tensile strain (CTS) (0.5Hz, 10% stretch) was applied for 30min. Total RNA was extracted, reverse transcribed, and analyzed by polymerase chain reaction (PCR) and real-time PCR. RUNX-2 overexpression and small interfering RNA (siRNA) targeting RUNX-2 were used to investigate the role of RUNX-2 in CTS-induced gene expression. The involvement of diverse mitogen-activated protein kinase (MAPK) pathways in the activation of RUNX-2, MMP-13 and ADAMTS-5 during CTS was examined by Western blotting. Results: CTS induced expression of RUNX-2, MMP-13, ADAMTS-4, -5, and -9. Overexpression of RUNX-2 up-regulated expression of MMP-13 and ADAMTS-5, whereas RUNX-2 siRNA resulted in significant down-regulation of mechanically-induced MMP-13 and ADAMTS-5 expression. CTS induced activation of p38 MAPK, and CTS induction of RUNX-2, MMP-13 and ADAMTS-5 mRNA was down-regulated by the selective p38 MAPK inhibitor SB203580 but not by the p44/42 MAPK inhibitor U0126, or the JNK MAPK inhibitor JNK inhibitor II. Conclusions: RUNX-2 might have a role as a key downstream mediator of p38's ability to regulate mechanical stress-induced MMP-13 and ADAMTS-5 expression.

Original languageEnglish
Pages (from-to)222-232
Number of pages11
JournalOsteoarthritis and Cartilage
Volume19
Issue number2
DOIs
Publication statusPublished - Feb 2011

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Mechanical Stress
Tensile strain
Chondrocytes
Matrix Metalloproteinases
Proteins
Protein Kinase Inhibitors
RNA
Polymerase chain reaction
p38 Mitogen-Activated Protein Kinases
Mitogen-Activated Protein Kinases
Small Interfering RNA
Chemical activation
Transcription factors
JNK Mitogen-Activated Protein Kinases
Gene expression
Real-Time Polymerase Chain Reaction
Transcription Factors
Down-Regulation
Western Blotting
Gene Expression

Keywords

  • ADAMTS
  • Aggrecanase
  • Chondrocyte
  • Mechanical stress
  • RUNX-2

ASJC Scopus subject areas

  • Biomedical Engineering
  • Orthopedics and Sports Medicine
  • Rheumatology

Cite this

@article{278cc575342648cbab228f57e5db8533,
title = "Regulation of mechanical stress-induced MMP-13 and ADAMTS-5 expression by RUNX-2 transcriptional factor in SW1353 chondrocyte-like cells",
abstract = "Objective: To investigate the mechanism of mechanical stress-induced expression and regulation of aggrecanases and examine the role of runt-related transcription factor 2 (RUNX-2) in chondrocyte-like cells. Methods: SW1353 cells were seeded onto stretch chambers at a concentration of 5×104cells/chamber, and a uni-axial cyclic tensile strain (CTS) (0.5Hz, 10{\%} stretch) was applied for 30min. Total RNA was extracted, reverse transcribed, and analyzed by polymerase chain reaction (PCR) and real-time PCR. RUNX-2 overexpression and small interfering RNA (siRNA) targeting RUNX-2 were used to investigate the role of RUNX-2 in CTS-induced gene expression. The involvement of diverse mitogen-activated protein kinase (MAPK) pathways in the activation of RUNX-2, MMP-13 and ADAMTS-5 during CTS was examined by Western blotting. Results: CTS induced expression of RUNX-2, MMP-13, ADAMTS-4, -5, and -9. Overexpression of RUNX-2 up-regulated expression of MMP-13 and ADAMTS-5, whereas RUNX-2 siRNA resulted in significant down-regulation of mechanically-induced MMP-13 and ADAMTS-5 expression. CTS induced activation of p38 MAPK, and CTS induction of RUNX-2, MMP-13 and ADAMTS-5 mRNA was down-regulated by the selective p38 MAPK inhibitor SB203580 but not by the p44/42 MAPK inhibitor U0126, or the JNK MAPK inhibitor JNK inhibitor II. Conclusions: RUNX-2 might have a role as a key downstream mediator of p38's ability to regulate mechanical stress-induced MMP-13 and ADAMTS-5 expression.",
keywords = "ADAMTS, Aggrecanase, Chondrocyte, Mechanical stress, RUNX-2",
author = "Tomonori Tetsunaga and Keiichiro Nishida and Takayuki Furumatsu and Keiji Naruse and Satoshi Hirohata and A. Yoshida and Taichi Saitou and Toshihumi Ozaki",
year = "2011",
month = "2",
doi = "10.1016/j.joca.2010.11.004",
language = "English",
volume = "19",
pages = "222--232",
journal = "Osteoarthritis and Cartilage",
issn = "1063-4584",
publisher = "W.B. Saunders Ltd",
number = "2",

}

TY - JOUR

T1 - Regulation of mechanical stress-induced MMP-13 and ADAMTS-5 expression by RUNX-2 transcriptional factor in SW1353 chondrocyte-like cells

AU - Tetsunaga, Tomonori

AU - Nishida, Keiichiro

AU - Furumatsu, Takayuki

AU - Naruse, Keiji

AU - Hirohata, Satoshi

AU - Yoshida, A.

AU - Saitou, Taichi

AU - Ozaki, Toshihumi

PY - 2011/2

Y1 - 2011/2

N2 - Objective: To investigate the mechanism of mechanical stress-induced expression and regulation of aggrecanases and examine the role of runt-related transcription factor 2 (RUNX-2) in chondrocyte-like cells. Methods: SW1353 cells were seeded onto stretch chambers at a concentration of 5×104cells/chamber, and a uni-axial cyclic tensile strain (CTS) (0.5Hz, 10% stretch) was applied for 30min. Total RNA was extracted, reverse transcribed, and analyzed by polymerase chain reaction (PCR) and real-time PCR. RUNX-2 overexpression and small interfering RNA (siRNA) targeting RUNX-2 were used to investigate the role of RUNX-2 in CTS-induced gene expression. The involvement of diverse mitogen-activated protein kinase (MAPK) pathways in the activation of RUNX-2, MMP-13 and ADAMTS-5 during CTS was examined by Western blotting. Results: CTS induced expression of RUNX-2, MMP-13, ADAMTS-4, -5, and -9. Overexpression of RUNX-2 up-regulated expression of MMP-13 and ADAMTS-5, whereas RUNX-2 siRNA resulted in significant down-regulation of mechanically-induced MMP-13 and ADAMTS-5 expression. CTS induced activation of p38 MAPK, and CTS induction of RUNX-2, MMP-13 and ADAMTS-5 mRNA was down-regulated by the selective p38 MAPK inhibitor SB203580 but not by the p44/42 MAPK inhibitor U0126, or the JNK MAPK inhibitor JNK inhibitor II. Conclusions: RUNX-2 might have a role as a key downstream mediator of p38's ability to regulate mechanical stress-induced MMP-13 and ADAMTS-5 expression.

AB - Objective: To investigate the mechanism of mechanical stress-induced expression and regulation of aggrecanases and examine the role of runt-related transcription factor 2 (RUNX-2) in chondrocyte-like cells. Methods: SW1353 cells were seeded onto stretch chambers at a concentration of 5×104cells/chamber, and a uni-axial cyclic tensile strain (CTS) (0.5Hz, 10% stretch) was applied for 30min. Total RNA was extracted, reverse transcribed, and analyzed by polymerase chain reaction (PCR) and real-time PCR. RUNX-2 overexpression and small interfering RNA (siRNA) targeting RUNX-2 were used to investigate the role of RUNX-2 in CTS-induced gene expression. The involvement of diverse mitogen-activated protein kinase (MAPK) pathways in the activation of RUNX-2, MMP-13 and ADAMTS-5 during CTS was examined by Western blotting. Results: CTS induced expression of RUNX-2, MMP-13, ADAMTS-4, -5, and -9. Overexpression of RUNX-2 up-regulated expression of MMP-13 and ADAMTS-5, whereas RUNX-2 siRNA resulted in significant down-regulation of mechanically-induced MMP-13 and ADAMTS-5 expression. CTS induced activation of p38 MAPK, and CTS induction of RUNX-2, MMP-13 and ADAMTS-5 mRNA was down-regulated by the selective p38 MAPK inhibitor SB203580 but not by the p44/42 MAPK inhibitor U0126, or the JNK MAPK inhibitor JNK inhibitor II. Conclusions: RUNX-2 might have a role as a key downstream mediator of p38's ability to regulate mechanical stress-induced MMP-13 and ADAMTS-5 expression.

KW - ADAMTS

KW - Aggrecanase

KW - Chondrocyte

KW - Mechanical stress

KW - RUNX-2

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U2 - 10.1016/j.joca.2010.11.004

DO - 10.1016/j.joca.2010.11.004

M3 - Article

VL - 19

SP - 222

EP - 232

JO - Osteoarthritis and Cartilage

JF - Osteoarthritis and Cartilage

SN - 1063-4584

IS - 2

ER -