Journal of Investigative Dermatology Home


Published:October 22, 2021DOI:


      Ultraviolet B (UVB) radiation directly damages DNA, increases reactive oxygen species (ROS) and nitric oxide (NO) release, and promotes inflammation leading to genomic instability and cell death. Nicotinamide (NAM) is the precursor of NAD, essential for cell energy production and DNA damage repair. NAM protects HaCat cells from UV-induced impairment; however, little is known about its effects on human primary keratinocytes (HPKs) and those isolated from field cancerization (FC-HPKs). We examined the role of NAM against UV-induced oxidative stress damages in FC-HPKs, isolated from precancerous lesions and skin cancers, and in normal epidermal keratinocytes (NHEK). Cells were treated for 18, 24, and 48 hours with NAM (5, 25, and 50 μM) before UVB irradiation. FC-HPK showed 4-fold higher basal ROS levels, comparing with NHEK; NAM downregulated ROS production only in irradiated FC-HPK, that showed a greater sensibility to UV rays. UV exposure increased OGG1, iNOS and IL-1β expression, an effect counteracted by NAM pre-treatment. Intracellular NO production and DNA damages were inhibited by NAM exposure before irradiation. Collectively, our findings indicate that pre-treatment with 25 μM NAM 24h before UVB irradiation effectively prevents oxidative stress formation, DNA damages, and inflammation in both NHEKs and FC-HPKs.

      Graphical abstract

      Key words


      ultraviolet ((UV)), reactive oxygen species ((ROS)), nitric oxide ((NO)), nicotinamide ((NAM)), human primary keratinocytes ((HPKs)), field cancerization human primary keratinocytes ((FC-HPKs)), Non-Melanoma Skin Cancers ((NMSCs)), 8-oxoguanine DNA glycosylase 1 ((OGG1)), inducible nitric oxide synthase ((iNOS)), cyclobutane pyrimidine dimers ((CPDs)), superoxide dismutase 1 ((SOD1)), 8-oxo-7,8-dihydro-2’-deoxyguanosine ((8oxoG)), DNA repair enzyme poly-ADP-ribose polymerase ((PARP)), cutaneous melanoma ((CM)), actinic keratoses ((AKs)), BCC ((basal cell carcinoma)), SCC ((squamous cell carcinoma))
      To read this article in full you will need to make a payment
      Purchase one-time access
      Society Members (SID/ESDR), remember to log in for access.
      Subscribe to Journal of Investigative Dermatology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Barthelemy H.
        • Chouvet B.
        • Cambazard F.
        Skin and mucosal manifestations in vitamin deficiency.
        J Am Acad Dermatol. 1986; 15: 1263-1274
        • Bierman J.C.
        • Laughlin T.
        • Tamura M.
        • et al.
        Niacinamide mitigates SASP-related inflammation induced by environmental stressors in human epidermal keratinocytes and skin [published online ahead of print, 2020 Jul 13].
        Int J Cosmet Sci. 2020; (10.1111/ics.12651. doi:10.1111/ics.12651)
        • Black A.T.
        • Gray J.P.
        • Shakarjian M.P.
        • Laskin D.L.
        • Heck D.E.
        • Laskin J.D.
        Distinct effects of ultraviolet B light on antioxidant expression in undifferentiated and differentiated mouse keratinocytes.
        Carcinogenesis. 2008; 29: 219-225
        • Bogan K.L.
        • Brenner C.
        Nicotinic acid, nicotinamide, and nicotinamide riboside: a molecular evaluation of NAD+ precursor vitamins in human nutrition.
        Annu Rev Nutr. 2008; 28: 115-130
        • Camillo L.
        • Gironi L.C.
        • Savoia P.
        Alpha-tocopherol promotes metabolic activity and cell migration in human primary keratinocytes and fibroblasts from skin cancer patients.
        G Ital Dermatol Venereol. 2018; (10.23736/S0392-0488.18.06075-3. doi:10.23736/S0392-0488.18.06075)
        • Chang H.R.
        • Tsao D.A.
        • Wang S.R.
        • Yu H.S.
        Expression of nitric oxide synthases in keratinocytes after UVB irradiation.
        Arch Dermatol Res. 2003 Dec; 295 (Epub 2003 Nov 13): 293-296
        • Chen A.C.
        • Damian D.L.
        Nicotinamide and the skin.
        Australas J Dermatol. 2014; 55 (Epub 2003 Nov 13): 169-175
        • Chen A.C.
        • Martin A.J.
        • Choy B.
        • Fernández-Peñas P.
        • Dalziell R.A.
        • McKenzie C.A.
        • et al.
        A Phase 3 Randomized Trial of Nicotinamide for Skin-Cancer Chemoprevention.
        N Engl J Med. 2015 22; 373: 1618-1626
        • de Berker D.
        • McGregor J.M.
        • Mohd Mustapa M.F.
        • Exton L.S.
        • Hughes B.R.
        British Association of Dermatologists' guidelines for the care of patients with actinic keratosis 2017.
        Br J Dermatol. 2017; 176: 20-43
        • de Jager T.L.
        • Cockrell A.E.
        • Du Plessis S.S.
        Ultraviolet Light Induced Generation of Reactive Oxygen Species.
        Adv Exp Med Biol. 2017; 996: 15-23
        • Fang L.
        • Neutzner A.
        • Turtschi S.
        • Flammer J.
        • Mozaffarieh M.
        Comet assay as an indirect measure of systemic oxidative stress.
        J Vis Exp. 2015 May 22; e52763
        • Fleming P.
        • Zhou S.
        • Bobotsis R.
        • Lynde C.
        Comparison of the Treatment Guidelines for Actinic Keratosis: A Critical Appraisal and Review.
        J Cutan Med Surg. 2017; 21: 408-417
        • Hageman G.J.
        • Stierum R.H.
        Niacin, poly(ADP-ribose) polymerase-1 and genomic stability.
        Mutat Res. 2001 Apr 18; 475: 45-56
        • Ichihashi M.
        • Ueda M.
        • Budiyanto A.
        • et al.
        UV-induced skin damage.
        Toxicology. 2003; 189: 21-39
        • Ikehata H.
        • Ono T.
        The mechanisms of UV mutagenesis.
        J Radiat Res. 2011; 52: 115-125
        • Jin G.H.
        • Liu Y.
        • Jin S.Z.
        • Liu X.D.
        • Liu S.Z.
        UVB induced oxidative stress in human keratinocytes and protective effect of antioxidant agents.
        Radiat Environ Biophys. 2007 Mar; 46: 61-68
        • Kabir Y.
        • Seidel R.
        • Mcknight B.
        • Moy R.
        DNA repair enzymes: an important role in skin cancer prevention and reversal of photodamage--a review of the literature.
        J Drugs Dermatol. 2015 Mar; 14: 297-303
        • Kirby J.S.
        • Scharnitz T.
        • Seiverling E.V.
        • Ahrns H.
        • Ferguson S.
        Actinic Keratosis Clinical Practice Guidelines: An Appraisal of Quality.
        Dermatol Res Pract. 2015; 2015: 456071
        • Lorsch J.R.
        • Collins F.S.
        • Lippincott-Schwartz J.
        Cell Biology. Fixing problems with cell lines.
        Science. 2014 Dec 19; 346: 1452-1453
        • Monfrecola G.
        • Gaudiello F.
        • Cirillo T.
        • Fabbrocini G.
        • Balato A.
        • Lembo S.
        Nicotinamide downregulates gene expression of interleukin-6, interleukin-10, monocyte chemoattractant protein-1, and tumour necrosis factor-α gene expression in HaCaT keratinocytes after ultraviolet B irradiation.
        Clin Exp Dermatol. 2013; 38: 185-188
        • Moscarella E.
        • Di Brizzi E.V.
        • Casari A.
        • De Giorgi V.
        • Di Meo N.
        • Fargnoli M.C.
        • et al.
        Italian expert consensus paper on the management of patients with actinic keratoses.
        Dermatol Ther. 2020 Jul 10; e13992
        • Nishisgori C.
        Current concept of photocarcinogenesis.
        Photochem Photobiol Sci. 2015 Sep 26; 14 (Epub 2015 Jul 15): 1713-1721
        • Park J.
        • Halliday G.M.
        • Surjana D.
        • Damian D.L.
        Nicotinamide prevents ultraviolet radiation-induced cellular energy loss.
        Photochem Photobiol. 2010 Jul-Aug; 86 (1097.2010.00746.x. Epub 2010 May 13): 942-948
        • Pelle E.
        • Huang X.
        • Mammone T.
        • Marenus K.
        • Maes D.
        • Frenkel K.
        Ultraviolet-B-induced oxidative DNA base damage in primary normal human epidermal keratinocytes and inhibition by a hydroxyl radical scavenger.
        J Invest Dermatol. 2003; 121: 177-183
        • Pillai S.
        • Oresajo C.
        • Hayward J.
        Ultraviolet radiation and skin aging: roles of reactive oxygen species, inflammation and protease activation, and strategies for prevention of inflammation-induced matrix degradation - a review.
        Int J Cosmet Sci. 2005; 27: 17-34
        • Savoia P.
        • Zavattaro E.
        • Cremona O.
        Chemoprevention of Skin Carcinomas in High-Risk Transplant Recipients.
        Curr Med Chem. 2018; 25: 687-697
        • Schade N.
        • Esser C.
        • Krutmann J.
        Ultraviolet B radiation-induced immunosuppression: molecular mechanisms and cellular alterations.
        Photochem Photobiol Sci. 2005; 4: 699-708
        • Seo S.J.
        • Choi H.G.
        • Chung H.J.
        • Hong C.K.
        Time course of expression of mRNA of inducible nitric oxide synthase and generation of nitric oxide by ultraviolet B in keratinocyte cell lines.
        Br J Dermatol. 2002; 147: 655-662
        • Snaidr V.A.
        • Damian D.L.
        • Halliday G.M.
        Nicotinamide for photoprotection and skin cancer chemoprevention: A review of efficacy and safety.
        Exp Dermatol. 2019; 28: 15-22
        • Stockfleth E.
        The importance of treating the field in actinic keratosis.
        J Eur Acad Dermatol Venereol. 2017; 31: 8-11
        • Surjana D.
        • Halliday G.M.
        • Martin A.J.
        • Moloney F.J.
        • Damian D.L.
        Oral nicotinamide reduces actinic keratoses in phase II double-blinded randomized controlled trials.
        J Invest Dermatol. 2012; 132 (10.1038/jid.2011.459. Epub 2012 Feb 2): 1497-1500
        • Surjana D.
        • Halliday G.M.
        • Damian D.L.
        Nicotinamide enhances repair of ultraviolet radiation-induced DNA damage in human keratinocytes and ex vivo skin.
        Carcinogenesis. 2013; 34: 1144-1149
        • Tan C.L.
        • Chin T.
        • Tan C.Y.R.
        • Rovito H.A.
        • Quek L.S.
        • Oblong J.E.
        • et al.
        Nicotinamide Metabolism Modulates the Proliferation/Differentiation Balance and Senescence of Human Primary Keratinocytes.
        J Invest Dermatol. 2019; 139 (e3): 1638-1647
        • Thompson B.C.
        • Halliday G.M.
        • Damian D.L.
        Nicotinamide enhances repair of arsenic and ultraviolet radiation-induced DNA damage in HaCaT keratinocytes and ex vivo human skin.
        PLoS One. 2015 Feb 6; 10 (10.1371/journal.pone.0117491. eCollection 2015)e0117491
        • Werner R.N.
        • Stockfleth E.
        • Connolly S.M.
        • Correia O.
        • Erdmann R.
        • Foley P.
        • et al.
        International League of Dermatological Societies; European Dermatology Forum. Evidence- and consensus-based (S3) Guidelines for the Treatment of Actinic Keratosis - International League of Dermatological Societies in cooperation with the European Dermatology Forum - Short version.
        J Eur Acad Dermatol Venereol. 2015; 29 (Epub 2015 Sep 14): 2069-2079
        • Willenbrink T.J.
        • Ruiz E.S.
        • Cornejo C.
        • Schmults C.D.
        • Arron S.
        • Jambusaria-Pahlajani A.
        Field Cancerization: Definition, epidemiology, risk factors, and outcomes.
        J Am Acad Dermatol. 2020; 6 (S0190-9622(20)30791-X)