Downregulation of a Rheumatoid Arthritis-Related Antigen (RA-A47) by ra-a47 Antisense Oligonucleotides Induces Inflammatory Factors in Chondrocytes

Takako Hattori, Harumi Kawaki, Satoshi Kubota, Yasutaka Yutani, Benoit De Crombrugghe, Klaus Von Der Mark, Masaharu Takigawa

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Previously we have shown that the expression of RA-A47 (rheumatoid arthritis-related antigen) which is identical to HSP47, a collagen-binding chaperon, is downregulated in chondrocytes by tumor necrosis factor α (TNFα). RA-A47 was also found on the surface of chondrocytes where it is recognized as an antigen in the serum of rheumatoid arthritis (RA) patients. Its translocation to the cell surface from endoplasmic reticulum membrane where it is normally located was also enhanced by TNFα. To understand the significance of RA-A47 downregulation in chondrocytes independent from other effects of TNFα, we used an antisense oligonucleotide approach and investigated the effect of this treatment on the expression of molecules related to matrix degradation and production of growth factors for chondrocytic, endothelial, and synovial cells. Here we show that treatment of rabbit chondrocyes and human chondrosarcoma cells HCS-2/8 by ra-a47 antisense S-oligonucleotides significantly reduced the expression of ra-a47 both at mRNA and protein level. Interestingly, this TNFα-independent RA-A47 downregulation was associated with a strong induction of matrix metalloproteinase (MMP)-9 mRNA and inducible NO synthase (iNOS) mRNA. The induction of active-type MMP-9 was further detected by gelatin zymography. Under the same conditions, the release of basic fibroblast growth factor (bFGF) and connective tissue growth factor (CTGF) from HCS-2/8 cells into the conditioned medium (CM) was strongly enhanced. These effects were not a result of TNFα upregulation, since the ra-a47 antisense oligonucleotide treatment did not enhance TNFα synthesis. These observations indicate that downregulation of RA-A47 induces TNFα-independent cartilage-degrading pathways involving iNOS and MMP-9. Furthermore, the stimulation of bFGF and CTGF release from chondrocytes may stimulate the proliferation of adjacent endothelial and/or synovial cells.

Original languageEnglish
Pages (from-to)94-102
Number of pages9
JournalJournal of Cellular Physiology
Volume197
Issue number1
DOIs
Publication statusPublished - Oct 2003

Fingerprint

Antisense Oligonucleotides
Chondrocytes
Rheumatoid Arthritis
Down-Regulation
Tumor Necrosis Factor-alpha
Antigens
Matrix Metalloproteinase 9
Connective Tissue Growth Factor
Fibroblast Growth Factor 2
Nitric Oxide Synthase
Messenger RNA
Endothelial Growth Factors
Chondrosarcoma
Cartilage
Gelatin
Conditioned Culture Medium
Oligonucleotides
Endoplasmic Reticulum
Intercellular Signaling Peptides and Proteins
Up-Regulation

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Downregulation of a Rheumatoid Arthritis-Related Antigen (RA-A47) by ra-a47 Antisense Oligonucleotides Induces Inflammatory Factors in Chondrocytes. / Hattori, Takako; Kawaki, Harumi; Kubota, Satoshi; Yutani, Yasutaka; De Crombrugghe, Benoit; Von Der Mark, Klaus; Takigawa, Masaharu.

In: Journal of Cellular Physiology, Vol. 197, No. 1, 10.2003, p. 94-102.

Research output: Contribution to journalArticle

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abstract = "Previously we have shown that the expression of RA-A47 (rheumatoid arthritis-related antigen) which is identical to HSP47, a collagen-binding chaperon, is downregulated in chondrocytes by tumor necrosis factor α (TNFα). RA-A47 was also found on the surface of chondrocytes where it is recognized as an antigen in the serum of rheumatoid arthritis (RA) patients. Its translocation to the cell surface from endoplasmic reticulum membrane where it is normally located was also enhanced by TNFα. To understand the significance of RA-A47 downregulation in chondrocytes independent from other effects of TNFα, we used an antisense oligonucleotide approach and investigated the effect of this treatment on the expression of molecules related to matrix degradation and production of growth factors for chondrocytic, endothelial, and synovial cells. Here we show that treatment of rabbit chondrocyes and human chondrosarcoma cells HCS-2/8 by ra-a47 antisense S-oligonucleotides significantly reduced the expression of ra-a47 both at mRNA and protein level. Interestingly, this TNFα-independent RA-A47 downregulation was associated with a strong induction of matrix metalloproteinase (MMP)-9 mRNA and inducible NO synthase (iNOS) mRNA. The induction of active-type MMP-9 was further detected by gelatin zymography. Under the same conditions, the release of basic fibroblast growth factor (bFGF) and connective tissue growth factor (CTGF) from HCS-2/8 cells into the conditioned medium (CM) was strongly enhanced. These effects were not a result of TNFα upregulation, since the ra-a47 antisense oligonucleotide treatment did not enhance TNFα synthesis. These observations indicate that downregulation of RA-A47 induces TNFα-independent cartilage-degrading pathways involving iNOS and MMP-9. Furthermore, the stimulation of bFGF and CTGF release from chondrocytes may stimulate the proliferation of adjacent endothelial and/or synovial cells.",
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AU - Kubota, Satoshi

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AU - Takigawa, Masaharu

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