International Journal of Oral Biology

The Korean Academy of Oral Biology (대한구강생물학회)
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Thrombospondins Mediate the Adhesion of Osteoblast to Extracelluar Matrix

  • Lim, Dong-Jin (Dept. of Pharmacology and Dental Therapeutics, Chonnam National University) ;
  • Bae, In-Ho (Dept. of Pharmacology and Dental Therapeutics, Chonnam National University) ;
  • Jeong, Byung-Chul (Dept. of Pharmacology and Dental Therapeutics, Chonnam National University) ;
  • Kim, Sun-Hun (Dental Science Research Institute and BK21 Project for Dentistry, Chonnam National University) ;
  • Park, Bae-Keun (Biomedical Research Center, Ulsan University Hospital) ;
  • Kang, In-Chul (Dental Science Research Institute and BK21 Project for Dentistry, Chonnam National University) ;
  • Lee, Shee-Eun (Dept. of Pharmacology and Dental Therapeutics, Chonnam National University) ;
  • Song, Sang-Hun (Department of Oral & Maxillofacial Surgery, College of Medicine, Hallym Univ. Medical center) ;
  • Koh, Jeong-Tae (Dept. of Pharmacology and Dental Therapeutics, Chonnam National University)
  • Published : 2008.09.30
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Abstract

Thrombospondins (TSP-1, TSP-2) are secretory extracellular glycoproteins that are involved in a variety of physiological processes such as tumor cell adhesion, invasion, and metastasis. The present study was undertaken to elucidate the involvement of thrombospondins in the adhesion of osteoblast-like cells using the TSP-1 or TSP-2 antisense MG63 and MC3T3-E1 cell lines. For downregulation of TSPs expression, we prepared antisense constructs for TSP-1 and TSP-2 using the pREP4 an episomal mammalian expression vector, which be able to produce the specific antisense oligonucleotides around chromosome. MG63 and MC3T3-E1 osteoblast-like cells were transfected with the antisense constructs and nonliposomal Fugene 6, and then selected under hygromycin B (50 ${\mu}g/ml$) treatment for 2 weeks. Western blot analysis revealed that expression of the TSP proteins was downregulated in the antisense cell lines. The cell adhesion assay showed that adhesive properties of TSP-1 and TSP-2 antisense MG63 cells on the polystyrene culture plate were reduced to 17% and 21% of the control cells, respectively, and those of the TSP-1 and TSP-2 antisense MC3T3-E1 cells also decreased to 19% and 27% of control, respectively. Adhesion of TSP-1 and TSP-2 antisense MC3T3-E1 cells on Type I collagen-coated culture plate decreased to 27% and 76%, respectively. These results indicate that TSP-1 and TSP-2 proteins may have an important role in adhesion of osteoblast-like cells to extracellular matrix.

Keywords

References

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