Inhibition of endogenous expression of connective tissue growth factor by its antisense oligonucleotide and antisense RNA suppresses proliferation and migration of vascular endothelial cells

Tsuyoshi Shimo, Tohru Nakanishi, Yusuke Kimura, Takashi Nishida, Kiyoto Ishizeki, Tomohiro Matsumura, Masaharu Takigawa

Research output: Contribution to journalArticle

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Abstract

Previously, we cloned an mRNA predominantly expressed in hypertrophic chondrocytes by differential display-PCR from a human chondrosarcoma-derived chondrocytic cell line (HCS-2/8) that is identical to that of connective tissue growth factor (CTGF). In the present study, we investigated the roles of CTGF in the proliferation and migration of vascular endothelial cells using its antisense oligonucleotide and antisense RNA, because angiogenesis into the hypertrophic zone of cartilage occurs at the final step of endochondral ossification. Immunohistochemical and immunofluorescence techniques revealed that not only hypertrophic chondrocytes but also endothelial cells in the cost-chondral junctions of mouse ribs were stained with an anti-CTGF antibody in vivo. Northern blot analysis revealed that CTGF was strongly expressed in chondrocytic cells as well as bovine aorta endothelial (BAE) cells in culture, but not in other types of cells such as osteoblastic: cells. Its expression in BAE cells was greater in the growing phase than in the confluent phase. When one-half of a monolayer of a confluent culture of BAE cells had been peeled off, only the cells proliferating and extending into the vacant area were stained with the anti-CTGF antibody. The addition of an antisense oligonucleotide inhibited the proliferation and extension of the BAE cells into the vacant area. The antisense oligonucleotide also inhibited the proliferation of BAE cells in the rapidly proliferating phase. In a Boyden chamber assay, pretreatment with the antisense oligonucleotide markedly inhibited the migration of BAE cells. Furthermore, the abilities to proliferate and migrate of BAE cells, which were stably transfected with expression vectors that generate the antisense RNA of CTGF cDNA, were markedly lower than those of the control. These findings suggest that endogenous CTGF expression is involved in the proliferation and migration of BAE cells.

Original languageEnglish
Pages (from-to)130-140
Number of pages11
JournalJournal of Biochemistry
Volume124
Issue number1
Publication statusPublished - Jul 1998

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Connective Tissue Growth Factor
Antisense RNA
Antisense Oligonucleotides
Endothelial cells
Endothelial Cells
Aorta
Chondrocytes
Cell culture
Cartilage
Chondrosarcoma
Antibodies
Ribs
Osteogenesis
Northern Blotting
Fluorescent Antibody Technique
Monolayers
Assays
Complementary DNA
Cell Culture Techniques
Display devices

Keywords

  • (BAE) cells
  • Angiogenesis
  • Antisense oligonucleotide
  • Antisense RNA
  • Bovine aorta endothelial
  • Connective tissue growth factor (CTGF)

ASJC Scopus subject areas

  • Biochemistry

Cite this

Inhibition of endogenous expression of connective tissue growth factor by its antisense oligonucleotide and antisense RNA suppresses proliferation and migration of vascular endothelial cells. / Shimo, Tsuyoshi; Nakanishi, Tohru; Kimura, Yusuke; Nishida, Takashi; Ishizeki, Kiyoto; Matsumura, Tomohiro; Takigawa, Masaharu.

In: Journal of Biochemistry, Vol. 124, No. 1, 07.1998, p. 130-140.

Research output: Contribution to journalArticle

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