Effects of CTGF/Hcs24, a product of a hypertrophic chondrocyte-specific gene, on the proliferation and differentiation of chondrocytes in culture

Tohru Nakanishi, Takashi Nishida, Tsuyoshi Shimo, Kappei Kobayashi, Toshikazu Kubo, Takuya Tamatani, Katsunari Tezuka, Masaharu Takigawa

Research output: Contribution to journalArticle

218 Citations (Scopus)

Abstract

Recently, we cloned a messenger RNA (mRNA) predominantly expressed in chondrocytes from a human chondrosarcoma-derived chondrocytic cell line, HCS-2/8, by differential display PCR and found that its gene, named hcs24, was identical with that of connective tissue growth factor (CTGF). Here we investigated CTGF/Hcs24 function in the chondrocytic cell line HCS-2/8 and rabbit growth cartilage (RGC) cells. HCS-2/8 cells transfected with recombinant adenoviruses that generate CTGF/Hcs24 sense RNA (mRNA) proliferated more rapidly than HCS-2/8 cells transfected with control adenoviruses. HCS-2/8 cells transfected with recombinant adenoviruses that generate CTGF/Hcs24 sense RNA expressed more mRNA of aggrecan and type X collagen than the control cells. To elucidate the direct action of CTGF/Hcs24 on the cells, we transfected HeLa cells with CTGF/Hcs24 expression vectors, obtained stable transfectants, and purified recombinant CTGF/Hcs24 protein from conditioned medium of the transfectants. The recombinant CTGF/Hcs24 effectively promoted the proliferation of HCS-2/8 cells and RGC cells in a dose-dependent manner and also dose dependently increased proteoglycan synthesis in these cells. In addition, these stimulatory effects of CTGF/Hcs24 were neutralized by the addition of anti-CTGF antibodies. Furthermore, the recombinant CTGF/Hcs24 effectively increased alkaline phosphatase activity in RGC cells in culture. Moreover, RT-PCR analysis revealed that the recombinant CTGF/Hcs24 stimulated gene expression of aggrecan and collagen types II and X in RGC cells in culture. These results indicate that CTGF/Hcs24 directly promotes the proliferation and differentiation of chondrocytes.

Original languageEnglish
Pages (from-to)264-273
Number of pages10
JournalEndocrinology
Volume141
Issue number1
DOIs
Publication statusPublished - 2000

Fingerprint

Connective Tissue Growth Factor
Chondrocytes
Genes
Cartilage
Collagen Type X
Adenoviridae
Rabbits
Aggrecans
Growth
Messenger RNA
Cell Culture Techniques
RNA
Cell Line
Polymerase Chain Reaction
Chondrosarcoma
Collagen Type II
Proteoglycans
Conditioned Culture Medium
HeLa Cells
Alkaline Phosphatase

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Effects of CTGF/Hcs24, a product of a hypertrophic chondrocyte-specific gene, on the proliferation and differentiation of chondrocytes in culture. / Nakanishi, Tohru; Nishida, Takashi; Shimo, Tsuyoshi; Kobayashi, Kappei; Kubo, Toshikazu; Tamatani, Takuya; Tezuka, Katsunari; Takigawa, Masaharu.

In: Endocrinology, Vol. 141, No. 1, 2000, p. 264-273.

Research output: Contribution to journalArticle

Nakanishi, Tohru ; Nishida, Takashi ; Shimo, Tsuyoshi ; Kobayashi, Kappei ; Kubo, Toshikazu ; Tamatani, Takuya ; Tezuka, Katsunari ; Takigawa, Masaharu. / Effects of CTGF/Hcs24, a product of a hypertrophic chondrocyte-specific gene, on the proliferation and differentiation of chondrocytes in culture. In: Endocrinology. 2000 ; Vol. 141, No. 1. pp. 264-273.
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