TGF-β is not involved in early phase growth inhibition of keratinocytes by 1α,25(OH) 2vitamin D 3

Yuji Shirakata, Hikaru Ueno, Yasushi Hanakawa, Kenji Kameda, Kenshi Yamasaki, Sho Tokumaru, Yoko Yahata, Mikiko Tohyama, Koji Sayama, Koji Hashimoto

Research output: Contribution to journalArticlepeer-review

21 Citations (Scopus)

Abstract

It has been proposed that transforming growth factor-β (TGF-β) is involved in the growth inhibition of normal human epidermal keratinocytes (NHEK) by 1α,25-dihydoxyvitamin D 3 (1α,25(OH) 2D 3), although this is still controversial because of the difficulty in blocking TGF-β activity completely. To determine whether TGF-β is involved in early phase growth inhibition by 1α,25(OH) 2D 3. TGF-β mRNA was detected by ribonuclease protection assay (RPA), and biological active TGF-β was determined by a luciferase reporter assay. To block intrinsic TGF-β activity completely, we constructed an adenovirus vector expressing a truncated TGF-β type II receptor with a dominant negative effect (AdexTβTR) that blocks TGF-β signal transduction. 1α,25(OH) 2D 3 slightly upregulated TGF-β1 and TGF-β2 after 24 h according to an RPA and a luciferase reporter assay, however growth inhibition by 1α,25(OH) 2D 3 occurred at 6 h. The addition of 10 -6 M of 1α,25(OH) 2D 3 to NHEK infected with AdexTβTR or AdexLacZ (control vector) reduced DNA synthesis to 59.3 and 62.2% at 6 h, respectively. There was no significant difference in cell number after a 3-day incubation with AdexTβTR or AdexLacZ-infected cells treated with 1α,25(OH) 2D 3. Since 1α,25(OH) 2D 3 rapidly inhibits NHEK growth regardless of the prevention of TGF-β signal transduction, TGF-β is not involved in early phase growth inhibition by 1α,25(OH) 2D 3.

Original languageEnglish
Pages (from-to)41-50
Number of pages10
JournalJournal of dermatological science
Volume36
Issue number1
DOIs
Publication statusPublished - 2004 Oct 1
Externally publishedYes

Keywords

  • 1α,25(OH) D
  • Adenovirus vector
  • Dominant negative TGF-β receptor
  • Growth inhibition
  • Keratinocyte
  • Luciferase reporter assay
  • RNase protection assay
  • TGF-β

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Dermatology

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