Synergistic effects of acitretin and narrow-band UVB on inducing the expression of heparin-binding epidermal-growth-factor-like growth factor in normal human keratinocytes
Abstract
It has been reported that heparin-binding epidermal-growth-factor-like growth factor (HB-EGF) plays an important role in regulating keratinocyte growth after retinoic acid treatment. In clinic, there are synergistic effects of retinoic acid plus ultraviolet (UVB) on improving efficacy and reducing the side effects in treating psoriasis, while the mechanism is not fully clear. Therefore, this study was taken to examine the alteration of cell proliferation and HB-EGF expression in cultured normal human keratinocytes after acitretin and/or narrow-band UVB (NB-UVB) exposure. After 12 h incubation with acitretin (0.1–1 μmol/L) and/or following NB-UVB (50–100 mJ/cm2) irradiation in normal human keratinocytes, the proliferation potency of the cells was evaluated by MTT colorimetric assay, and HB-EGF protein and mRNA was detected by cytoimmunochemistry and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR), respectively. Our results revealed 1 μmol/L acitretin combined with 100 mJ/cm2 NB-UVB showed strongest inhibition of keratinocyte proliferation and highest induction of HB-EGF mRNA expression (19.7% and 8.6-fold, respectively), the others were 1 μmol/L acitretin (14.4%, 7.1-fold), 0.1 μmol/L acitretin (10.2%, 3.2-fold), 100 mJ/cm2 NB-UVB (8.7%, 2.5-fold) and 50 mJ/cm2 NB-UVB (5.4%, 1.8-fold). Pattern of HB-EGF protein expression was similar to mRNA expression in keratinocytes upon above cultural conditions. Our results suggest acitretin or NB-UVB single treatment can dose-dependently inhibit keratinocyte proliferation and induce HB-EGF expression and there are synergistic effects when they are combined.
Keywords
Acitretin HB-EGF Keratinocyte UVBNotes
Acknowledgments
We thank Shengxiang Xiao and Zhenping Cao for their technical assistance. We also acknowledge Wei Wang for providing the foreskin from plastic surgery. This work was supported by the National Natural Science Foundation of China (No. 30371295 and No. 30400387).
References
- 1.Alibardi L (2003) Immunocytochemistry and keratinization in the epidermis of crocodilians. Zool Stud 42:346–356Google Scholar
- 2.Arakawa S, Hatano Y, Katagiri K, Terashi H, Fujiwara S (2006) Effects of ultraviolet B irradiation on the production of regulated upon activation normal T-cell expressed and secreted protein in cultured human epidermal keratinocytes. Arch Dermatol Res 297:377–380PubMedCrossRefGoogle Scholar
- 3.Aufiero BM, Talwar H, Young C, Krishnan M, Hatfield JS, Lee HK, Wong HK, Hamzavi I, Murakawa GJ (2006) Narrow-band UVB induces apoptosis in human keratinocytes. J Photochem Photobiol B 82:132–139PubMedCrossRefGoogle Scholar
- 4.Boudjelal M, Voorhees JJ, Fisher GJ (2002) Retinoid signaling is attenuated by proteasome-mediated degradation of retinoid receptors in human keratinocyte HaCaT cells. Exp Cell Res 274:130–137PubMedCrossRefGoogle Scholar
- 5.Iwamoto R, Mekada E (2000) Heparin-binding EGF-like growth factor: a juxtacrine growth factor. Cytokine Growth Factor Rev 11:335–344PubMedCrossRefGoogle Scholar
- 6.Kimura R, Iwamoto R, Mekada E (2005) Soluble form of heparin-binding EGF-like growth factor contributes to retinoic acid-induced epidermal hyperplasia. Cell Struct Funct 30:35–42PubMedCrossRefGoogle Scholar
- 7.Lewis DA, Hurwitz SA, Spandau DF (2003) UVB-induced apoptosis in normal human keratinocytes: role of the erbB receptor family. Exp Cell Res 284:316–327PubMedCrossRefGoogle Scholar
- 8.Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408PubMedCrossRefGoogle Scholar
- 9.Lowe NL, Prystowsky JH, Bourget T, Edelstein J, Nychay S, Armstrong R (1991) Acitretin plus UVB therapy for psoriasis. Comparisons with placebo plus UVB and acitretin alone. J Am Acad Dermatol 24:591–594PubMedCrossRefGoogle Scholar
- 10.Miyamoto S, Yagi H, YotsμMoto F, Kawarabayashi T, Mekada E (2006) Heparin-binding epidermal growth factor-like growth factor as a novel targeting molecule for cancer therapy. Cancer Sci 97:341–347PubMedCrossRefGoogle Scholar
- 11.Nanba D, Higashiyama S (2004) Dual intracellular signaling by proteolytic cleavage of membrane-anchored heparin-binding EGF-like growth factor. Cytokine Growth Factor Rev 15:13–19PubMedCrossRefGoogle Scholar
- 12.Piepkorn M, Predd H, Underwood R, Cook P (2003) Proliferation–differentiation relationships in the expression of heparin-binding epidermal growth factor-related factors and erbB receptors by normal and psoriatic human keratinocytes. Arch Dermatol Res 295:93–101PubMedCrossRefGoogle Scholar
- 13.Rittie L, Varani J, Kang S, Voorhees JJ, Fisher GJ (2006) Retinoid-induced epidermal hyperplasia is mediated by epidermal growth factor receptor activation via specific induction of its ligands heparin-binding EGF and amphiregulin in human skin in vivo. J Invest Dermatol 126:732–739PubMedCrossRefGoogle Scholar
- 14.Seqaert S, Garmyn M, Bouillon R, Degreef H (1995) Subpharmacological concentrations of retinoic acid attenuate the antiproliferative effect of calcitriol on cultured human keratinocytes. J Eur Acad Dermatol Venereol 5: S91CrossRefGoogle Scholar
- 15.Varani J, Zeigler M, Dame MK, Kang S, Fisher GJ, Voorhees JJ, Stoll SW, Elder JT (2001) Heparin-binding epidermal-growth-factor-like growth factor activation of keratinocyte ErbB receptors mediates epidermal hyperplasia, a prominent side-effect of retinoid therapy. J Invest Dermatol 117:1335–1341PubMedCrossRefGoogle Scholar
- 16.Yu WH, Woessner JF, McNeish JD, Stamenkovic I (2002) CD44 anchors the assembly of matrilysin/MMP-7 with heparin-binding epidermal growth factor precursor and ErbB4 and regulates female reproductive organ remodeling. Genes Dev 16:307–323PubMedCrossRefGoogle Scholar