Regulation of rat bone sialoprotein gene transcription by enamel matrix derivative

Emi Shimizu, Yu Nakajima, Naoko Kato, Youhei Nakayama, Ryoichiro Saito, Hiroshi Samoto, Yorimasa Ogata

Research output: Contribution to journalArticle

Abstract

Background: Enamel matrix derivative (EMD) has recently been developed for use as a periodontal regenerative treatment. While EMD is believed to induce regeneration of periodontal tissue, the precise mechanism is not known. Bone sialoprotein (BSP), an early phenotypic marker of osteoblast differentiation, has been implicated in the nucleation of hydroxyapatite during bone formation. In this study, we examined the ability of EMD to regulate BSP gene transcription in osteoblast-like cells. Methods: To determine the molecular basis of the transcriptional regulation of BSP gene transcription by EMD, we conducted Northern hybridization, transient transfection analyses with chimeric constructs of the rat BSP gene promoter linked to a luciferase reporter gene, and gel mobility shift assays. Results: Using the osteoblastic cell line ROS 17/2.8, we determined that BSP mRNA levels increased ∼2.8-fold by EMD. In transient transfection analyses, EMD (50 μg/ml, 12 hours) increased luciferase activities of pLUC4 (nt -425 to +60) and pLUC5 (nt -801 to +60), transfected into ROS 17/2.8 cells. Within the pLUC4 and 5, a homeodomain binding element (HOX) and a transforming growth factor (TGF)-β activation element (TAE) are present. Gel mobility shift assays with radiolabeled HOX and TAE ds-oligonucleotides revealed increased binding of nuclear proteins from EMD stimulated ROS 17/2.8 cells. Conclusion: These studies have, therefore, identified EMD response elements in the rat BSP gene promoter that may mediates the effects of EMD on BSP gene transcription.

Original languageEnglish (US)
Pages (from-to)260-267
Number of pages8
JournalJournal of Periodontology
Volume75
Issue number2
DOIs
StatePublished - Feb 2004

Fingerprint

Integrin-Binding Sialoprotein
Dental Enamel
Genes
Electrophoretic Mobility Shift Assay
Luciferases
Osteoblasts
Transfection
Gels
Nuclear Matrix-Associated Proteins
rat Spp1 protein
Differentiation Antigens
Response Elements
Transforming Growth Factors
Durapatite
Reporter Genes
Osteogenesis
Oligonucleotides
Regeneration
Cell Line

Keywords

  • Animal studies
  • Enamel matrix derivative
  • Genetic
  • Periodontal regeneration
  • Sialoproteins
  • Transcription

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Shimizu, E., Nakajima, Y., Kato, N., Nakayama, Y., Saito, R., Samoto, H., & Ogata, Y. (2004). Regulation of rat bone sialoprotein gene transcription by enamel matrix derivative. Journal of Periodontology, 75(2), 260-267. https://doi.org/10.1902/jop.2004.75.2.260

Regulation of rat bone sialoprotein gene transcription by enamel matrix derivative. / Shimizu, Emi; Nakajima, Yu; Kato, Naoko; Nakayama, Youhei; Saito, Ryoichiro; Samoto, Hiroshi; Ogata, Yorimasa.

In: Journal of Periodontology, Vol. 75, No. 2, 02.2004, p. 260-267.

Research output: Contribution to journalArticle

Shimizu, E, Nakajima, Y, Kato, N, Nakayama, Y, Saito, R, Samoto, H & Ogata, Y 2004, 'Regulation of rat bone sialoprotein gene transcription by enamel matrix derivative', Journal of Periodontology, vol. 75, no. 2, pp. 260-267. https://doi.org/10.1902/jop.2004.75.2.260
Shimizu, Emi ; Nakajima, Yu ; Kato, Naoko ; Nakayama, Youhei ; Saito, Ryoichiro ; Samoto, Hiroshi ; Ogata, Yorimasa. / Regulation of rat bone sialoprotein gene transcription by enamel matrix derivative. In: Journal of Periodontology. 2004 ; Vol. 75, No. 2. pp. 260-267.
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