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Shirakata, Y. et al.

Shirakata, Ueno, Hanakawa, Kameda, Yamasaki, Tokumaru, Yahata, Tohyama, Sayama, Hashimoto,

TGF-beta is not involved in early phase growth inhibition of keratinocytes by 1alpha,25(OH)2vitamin D3.

BACKGROUND: It has been proposed that transforming growth factor-beta (TGF-beta) is involved in the growth inhibition of normal human epidermal keratinocytes (NHEK) by 1alpha,25-dihydoxyvitamin D(3) (1alpha,25(OH)(2)D(3)), although this is still controversial because of the difficulty in blocking TGF-beta activity completely. OBJECTIVE: To determine whether TGF-beta is involved in early phase growth inhibition by 1alpha,25(OH)(2)D(3). METHODS: TGF-beta mRNA was detected by ribonuclease protection assay (RPA), and biological active TGF-beta was determined by a luciferase reporter assay. To block intrinsic TGF-beta activity completely, we constructed an adenovirus vector expressing a truncated TGF-beta type II receptor with a dominant negative effect (AdexTbetaTR) that blocks TGF-beta signal transduction. RESULTS: 1alpha,25(OH)(2)D(3) slightly upregulated TGF-beta1 and TGF-beta2 after 24 h according to an RPA and a luciferase reporter assay, however growth inhibition by 1alpha,25(OH)(2)D(3) occurred at 6 h. The addition of 10(-6) M of 1alpha,25(OH)(2)D(3) to NHEK infected with AdexTbetaTR 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 AdexTbetaTR or AdexLacZ-infected cells treated with 1alpha,25(OH)(2)D(3). CONCLUSION: Since 1alpha,25(OH)(2)D(3) rapidly inhibits NHEK growth regardless of the prevention of TGF-beta signal transduction, TGF-beta is not involved in early phase growth inhibition by 1alpha,25(OH)(2)D(3).[1]

References

TGF-beta is not involved in early phase growth inhibition of keratinocytes by 1alpha,25(OH)2vitamin D3. Shirakata, Y., Ueno, H., Hanakawa, Y., Kameda, K., Yamasaki, K., Tokumaru, S., Yahata, Y., Tohyama, M., Sayama, K., Hashimoto, K. J. Dermatol. Sci. (2004) [Pubmed]

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