The effects of phototherapy and melanocytes on keratinocytes

Tang,Luyan; Wu,Wenyu; Fu,Wenwen; Hu,Yao

doi:10.3892/etm.2018.5807

The effects of phototherapy and melanocytes on keratinocytes

Authors:
- Luyan Tang
- Wenyu Wu
- Wenwen Fu
- Yao Hu
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Affiliations: Department of Dermatology, Huashan Hospital of Fudan University, Shanghai 200040, P.R. China
Published online on: January 30, 2018 https://doi.org/10.3892/etm.2018.5807
Pages: 3459-3466

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Abstract

Phototherapy is widely used in the treatment of vitiligo. Previous studies have focused on the effects of ultraviolet (UV) radiation on melanocytes; however, the biological effects of phototherapy and melanocytes on keratinocytes remain to be elucidated. To investigate and assess the effects of clinically doses of broad band (BB)‑UVA, narrow band (NB)‑UVB and melanocytes on human keratinocytes in vitro, clinical doses of BB‑UVA or NB‑UVB radiation and human melanoma cell A375 co‑culture were performed as stress divisors to HaCaT cells. Cell proliferation, expression of protease‑activated receptor‑2 (PAR‑2) and nuclear factor E2‑related factor 2 mRNA, lipid peroxidation and intracellular antioxidant level of keratinocytes were analyzed. It was demonstrated that UV radiation inhibited the proliferation of cells apart from following exposure to low dose (1 J/cm2) UVA. Medium dose (5 J/cm2) UVA radiation had no adverse effects on lipid peroxidation and increased antioxidant levels in HaCaT cells. Medium (200 mJ/cm2) and high (400 mJ/cm2) doses of UVB radiation induced cellular damage due to increased lipid peroxidation as indicated by levels of malondialdehyde. Furthermore, A375 co‑culture treatment induced a similar effect on the lipid peroxidation of HaCaT as with low dose UVB radiation. Therefore, the results of the present study determined that clinical doses of BB‑UVA and NB‑UVB radiation had varying effects on proliferation and related protein levels in HaCaT cells. Co‑culture with A375 had similar effects as those of low dose UVA and UVB radiation, in which the PAR‑2 expression was significantly upregulated.

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1	Njoo MD, Spuls PI, Bos JD, Westerhof W and Bossuyt PM: Nonsurgical repigmentation therapies in vitiligo-Meta-analysis of the literature. Arch Dermatol. 134:1532–1540. 1998. View Article : Google Scholar : PubMed/NCBI
2	Ezzedine K, Eleftheriadou V, Whitton M and van Geel N: Vitiligo. Lancet. 386:74–84. 2015. View Article : Google Scholar : PubMed/NCBI
3	Chu ECY: Treatment of vitiligo: Medical treatment, phototherapy and surgical treatment. Hong Kong J Dermatol Venereol. 23:113–117. 2015.
4	Kanwar AJ, Dogra S, Parsad D and Kumar B: Narrow-band UVB for the treatment of vitiligo: An emerging effective and well-tolerated therapy. Int J Dermatol. 44:57–60. 2005. View Article : Google Scholar : PubMed/NCBI
5	Wu CS, Yu CL, Wu CS, Lan CCE and Yu HS: Narrow-band ultraviolet-B stimulates proliferation and migration of cultured melanocytes. Exp Dermatol. 13:755–763. 2004. View Article : Google Scholar : PubMed/NCBI
6	El Mofty M, Bosseila M, Mashaly HM, Gawdat H and Makaly H: Broadband ultraviolet A vs. psoralen ultraviolet A in the treatment of vitiligo: A randomized controlled trial. Clin Exp Dermatol. 38:830–835. 2013. View Article : Google Scholar : PubMed/NCBI
7	Huang XX, Bernerd F and Halliday GM: Ultraviolet a within sunlight induces mutations in the epidermal basal layer of engineered human skin. Am J Pathol. 174:1534–1543. 2009. View Article : Google Scholar : PubMed/NCBI
8	Yasui H and Sakurai H: Chemiluminescent detection and imaging of reactive oxygen species in live mouse skin exposed to UVA. Biochem Biophys Res Commun. 269:131–136. 2000. View Article : Google Scholar : PubMed/NCBI
9	He YY and Häder DP: UV-B-induced formation of reactive oxygen species and oxidative damage of the cyanobacterium Anabaena sp.: Protective effects of ascorbic acid and N-acetyl-L-cysteine. J Photochem Photobiol B. 66:115–124. 2002. View Article : Google Scholar : PubMed/NCBI
10	Swalwell H, Latimer J, Haywood RM and Birch-Machin MA: Investigating the role of melanin in UVA/UVB-and hydrogen peroxide-induced cellular and mitochondrial ROS production and mitochondrial DNA damage in human melanoma cells. Free Radic Biol Med. 52:626–634. 2012. View Article : Google Scholar : PubMed/NCBI
11	Pelle E, Mammone T, Maes D and Frenkel K: Keratinocytes act as a source of reactive oxygen species by transferring hydrogen peroxide to melanocytes. J Invest Dermatol. 124:793–797. 2005. View Article : Google Scholar : PubMed/NCBI
12	Sravani PV, Babu NK, Gopal KV, Rao GR, Rao AR, Moorthy B and Rao TR: Determination of oxidative stress in vitiligo by measuring superoxide dismutase and catalase levels in vitiliginous and non-vitiliginous skin. Indian J Dermatol Venereol Leprol. 75:268–271. 2009. View Article : Google Scholar : PubMed/NCBI
13	Chui DH, Tang W and Orkin SH: cDNA cloning of murine Nrf 2 gene, coding for a p45 NF-E2 related transcription factor. Biochem Biophys Res Commun. 209:40–46. 1995. View Article : Google Scholar : PubMed/NCBI
14	McMahon M, Itoh K, Yamamoto M and Hayes JD: Keap1-dependent proteasomal degradation of transcription factor Nrf2 contributes to the negative regulation of antioxidant response element-driven gene expression. J Biol Chem. 278:21592–21600. 2003. View Article : Google Scholar : PubMed/NCBI
15	Itoh K, Wakabayashi N, Katoh Y, Ishii T, Igarashi K, Engel JD and Yamamoto M: Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain. Genes Dev. 13:76–86. 1999. View Article : Google Scholar : PubMed/NCBI
16	Déry O, Corvera CU, Steinhoff M and Bunnett NW: Proteinase-activated receptors: Novel mechanisms of signaling by serine proteases. Am J Physiol. 274:C1429–C1452. 1998. View Article : Google Scholar : PubMed/NCBI
17	Moretti S, Nassini R, Prignano F, Pacini A, Materazzi S, Naldini A, Simoni A, Baroni G, Pellerito S, Filippi I, et al: Protease-activated receptor-2 downregulation is associated to vitiligo lesions. Pigment Cell Melanoma Res. 22:335–338. 2009. View Article : Google Scholar : PubMed/NCBI
18	Rattenholl A and Steinhoff M: Proteinase-activated receptor-2 in the skin: Receptor expression, activation and function during health and disease. Drug News Perspect. 21:369–381. 2008. View Article : Google Scholar : PubMed/NCBI
19	Hachem JP, Houben E, Crumrine D, Man MQ, Schurer N, Roelandt T, Choi EH, Uchida Y, Brown BE, Feingold KR and Elias PM: Serine protease signaling of epidermal permeability barrier homeostasis. J Invest Dermatol. 126:2074–2086. 2006. View Article : Google Scholar : PubMed/NCBI
20	Hachem JP, Man MQ, Crumrine D, Uchida Y, Brown BE, Rogiers V, Roseeuw D, Feingold KR and Elias PM: Sustained serine proteases activity by prolonged increase in pH leads to degradation of lipid processing enzymes and profound alterations of barrier function and stratum corneum integrity. J Invest Dermatol. 125:510–520. 2005. View Article : Google Scholar : PubMed/NCBI
21	Steinhoff M, Corvera CU, Thoma MS, Kong W, McAlpine BE, Caughey GH, Ansel JC and Bunnett NW: Proteinase-activated receptor-2 in human skin: Tissue distribution and activation of keratinocytes by mast cell tryptase. Exp Dermatol. 8:282–294. 1999. View Article : Google Scholar : PubMed/NCBI
22	Steinhoff M, Neisius U, Ikoma A, Fartasch M, Heyer G, Skov PS, Luger TA and Schmelz M: Proteinase-activated receptor-2 mediates itch: A novel pathway for pruritus in human skin. J Neurosci. 23:6176–6180. 2003.PubMed/NCBI
23	Scott G, Deng A, Rodriguez-Burford C, Seiberg M, Han R, Babiarz L, Grizzle W, Bell W and Pentland A: Protease-activated receptor 2, a receptor involved in melanosome transfer, is upregulated in human skin by ultraviolet irradiation. J Invest Dermatol. 117:1412–1420. 2001. View Article : Google Scholar : PubMed/NCBI
24	Sasaki M, Horikoshi T, Uchiwa H and Miyachi Y: Up-regulation of tyrosinase gene by nitric oxide in human melanocytes. Pigment Cell Res. 13:248–252. 2000. View Article : Google Scholar : PubMed/NCBI
25	Roméro-Graillet C, Aberdam E, Clément M, Ortonne JP and Ballotti R: Nitric oxide produced by ultraviolet-irradiated keratinocytes stimulates melanogenesis. J Clin Invest. 99:635–642. 1997. View Article : Google Scholar : PubMed/NCBI
26	El-Mofty M, Mostafa W, Youssef R, El-Fangary M, El-Ramly A, Mahgoub D, Fawzy M and El-Hawary M: BB-UVA vs. NB-UVB in the treatment of vitiligo: A randomized controlled clinical study (single blinded). Photodermatol Photoimmunol Photomed. 29:239–246. 2013. View Article : Google Scholar : PubMed/NCBI
27	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
28	Ohkawa H, Ohishi N and Yagi K: Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 95:351–358. 1979. View Article : Google Scholar : PubMed/NCBI
29	Plaa GL and Witschi H: Chemicals, drugs, and lipid peroxidation. Annu Rev Pharmacol Toxicol. 16:125–142. 1976. View Article : Google Scholar : PubMed/NCBI
30	Roelandts R: Photo (chemo) therapy for vitiligo. Photodermatol Photoimmunol Photomed. 19:1–4. 2003. View Article : Google Scholar : PubMed/NCBI
31	Wang HT, Choi B and Tang MS: Melanocytes are deficient in repair of oxidative DNA damage and UV-induced photoproducts. Proc Natl Acad Sci USA. 107:pp. 12180–12185. 2010; View Article : Google Scholar : PubMed/NCBI
32	Roméro-Graillet C, Aberdam E, Clément M, Ortonne JP and Ballotti R: Nitric oxide produced by ultraviolet-irradiated keratinocytes stimulates melanogenesis. J Clin Invest. 99:635–642. 1997. View Article : Google Scholar : PubMed/NCBI
33	He YY, Pi J, Huang JL, Diwan BA, Waalkes MP and Chignell CF: Chronic UVA irradiation of human HaCaT keratinocytes induces malignant transformation associated with acquired apoptotic resistance. Oncogene. 25:3680–3688. 2006. View Article : Google Scholar : PubMed/NCBI
34	He YY, Huang JL and Chignell CF: Delayed and sustained activation of extracellular signal-regulated kinase in human keratinocytes by UVA: Implications in carcinogenesis. J Biol Chem. 279:53867–53874. 2004. View Article : Google Scholar : PubMed/NCBI
35	Lehmann J, Pollet D, Peker S, Steinkraus V and Hoppe U: Kinetics of DNA strand breaks and protection by antioxidants in UVA- or UVB-irradiated HaCaT keratinocytes using the single cell gel electrophoresis assay. Mutat Res. 407:97–108. 1998. View Article : Google Scholar : PubMed/NCBI
36	Hirota A, Kawachi Y, Itoh K, Nakamura Y, Xu X, Banno T, Takahashi T, Yamamoto M and Otsuka F: Ultraviolet A irradiation induces NF-E2-related factor 2 activation in dermal fibroblasts: Protective role in UVA-induced apoptosis. J Invest Dermatol. 124:825–832. 2005. View Article : Google Scholar : PubMed/NCBI
37	Hseu YC, Chou CW, Senthil Kumar KJ, Fu KT, Wang HM, Hsu LS, Kuo YH, Wu CR, Chen SC and Yang HL: Ellagic acid protects human keratinocyte (HaCaT) cells against UVA-induced oxidative stress and apoptosis through the upregulation of the HO-1 and Nrf-2 antioxidant genes. Food Chem Toxicol. 50:1245–1255. 2012. View Article : Google Scholar : PubMed/NCBI
38	Kimura S, Warabi E, Yanagawa T, Ma D, Itoh K, Ishii Y, Kawachi Y and Ishii T: Essential role of Nrf2 in keratinocyte protection from UVA by quercetin. Biochem Biophys Res Commun. 387:109–114. 2009. View Article : Google Scholar : PubMed/NCBI
39	Marrot L, Jones C, Perez P and Meunier JR: The significance of Nrf2 pathway in (photo)-oxidative stress response in melanocytes and keratinocytes of the human epidermis. Pigment Cell Melanoma Res. 21:79–88. 2008. View Article : Google Scholar : PubMed/NCBI
40	Bhatnagar A, Kanwar A, Parsad D and De D: Comparison of systemic PUVA and NB-UVB in the treatment of vitiligo: An open prospective study. J Eur Acad Dermatol Venereol. 21:638–642. 2007.PubMed/NCBI
41	Armstrong BK and Kricker A: The epidemiology of UV induced skin cancer. J Photochem Photobiol B. 63:8–18. 2001. View Article : Google Scholar : PubMed/NCBI
42	Rhodes LE, Belgi G, Parslew R, McLoughlin L, Clough GF and Friedmann PS: Ultraviolet-B-induced erythema is mediated by nitric oxide and prostaglandin E 2 in combination. J Invest Dermatol. 117:880–885. 2001. View Article : Google Scholar : PubMed/NCBI
43	Brash DE, Ziegler A, Jonason AS, Simon JA, Kunala S and Leffell DJ: Sunlight and sunburn in human skin cancer: p53, apoptosis, and tumor promotion. J Investig Dermatol Symp Proc. 1:136–142. 1996.PubMed/NCBI
44	Faraonio R, Vergara P, Di Marzo D, Pierantoni MG, Napolitano M, Russo T and Cimino F: p53 suppresses the Nrf2-dependent transcription of antioxidant response genes. J Biol Chem. 281:39776–39784. 2006. View Article : Google Scholar : PubMed/NCBI
45	Rushworth SA, Zaitseva L, Murray MY, Shah NM, Bowles KM and MacEwan DJ: The high Nrf2 expression in human acute myeloid leukemia is driven by NF-κB and underlies its chemo-resistance. Blood. 120:5188–5198. 2012. View Article : Google Scholar : PubMed/NCBI
46	Joshi PG, Nair N, Begum G, Joshi NB, Sinkar VP and Vora S: Melanocyte-keratinocyte interaction induces calcium signalling and melanin transfer to keratinocytes. Pigment Cell Res. 20:380–384. 2007.PubMed/NCBI
47	Scott G, Deng A, Rodriguez-Burford C, Seiberg M, Han R, Babiarz L, Grizzle W, Bell W and Pentland A: Protease-activated receptor 2, a receptor involved in melanosome transfer, is upregulated in human skin by ultraviolet irradiation. J Invest Dermatol. 117:1412–1420. 2001. View Article : Google Scholar : PubMed/NCBI
48	Paine C, Sharlow E, Liebel F, Eisinger M, Shapiro S and Seiberg M: An alternative approach to depigmentation by soybean extracts via inhibition of the PAR-2 pathway. J Invest Dermatol. 116:587–595. 2001. View Article : Google Scholar : PubMed/NCBI
49	Lerner DJ, Chen M, Tram T and Coughlin SR: Agonist recognition by proteinase-activated receptor 2 and thrombin receptor. Importance of extracellular loop interactions for receptor function. J Biol Chem. 271:13943–13947. 1996. View Article : Google Scholar : PubMed/NCBI
50	Molino M, Barnathan ES, Numerof R, Clark J, Dreyer M, Cumashi A, Hoxie JA, Schechter N, Woolkalis M and Brass LF: Interactions of mast cell tryptase with thrombin receptors and PAR-2. J Biol Chem. 272:4043–4049. 1997. View Article : Google Scholar : PubMed/NCBI
51	Molino M, Woolkalis MJ, Reavey-Cantwell J, Praticó D, Andrade-Gordon P, Barnathan ES and Brass LF: Endothelial cell thrombin receptors and PAR-2. Two protease-activated receptors located in a single cellular environment. J Biol Chem. 272:11133–11141. 1997. View Article : Google Scholar : PubMed/NCBI
52	Scott G, Leopardi S, Parker L, Babiarz L, Seiberg M and Han R: The proteinase-activated receptor-2 mediates phagocytosis in a Rho-dependent manner in human keratinocytes. J Invest Dermatol. 121:529–541. 2003. View Article : Google Scholar : PubMed/NCBI