A comparative study of UV-induced cell signalling pathways in human keratinocyte-derived cell lines
Tóm tắt
Ultraviolet (UV) radiation can activate the p38 mitogen-activated protein kinase (MAPK), Jun N-terminal kinase (JNK) and nuclear factor-κB (NFκB) pathways in skin cells. HaCaT cells are widely used as a primary keratinocyte substitute to study these pathways. However, like most squamous cell carcinomas (SCCs), it contains a dysfunctional p53. It is unclear if HaCaT cells activate these signalling pathways similarly to SCC cells (Colo16) or to primary human epidermal keratinocytes (HEK). In this study, the UV activation (UVA, UVB, UVA + B, UVB + A) of p38 MAPK, JNK and NFκB pathways, and TNFα secretion by HEK, HaCaT and Colo16 cells were investigated. The signalling pathway activation was UV-type and dose-dependent with UVB + A radiation inducing a high p38 and JNK activation. HaCaT cells exhibited 2- to 4-fold higher activity of the p38 (771 % at 60 min) and JNK (794 % at 30 min) pathways following UVB + A radiation than did HEK cells (p38: 367 % at 15 min and JNK: 184 % at 30 min). While both HaCaT and Colo16 cells did not activate the NFκB pathway, Colo16 cells had a lower p38 and higher JNK activity than HaCaT cells. Irradiated HaCaT cells produced less TNFα (UVB: 3.5 pg/ml), while HEK cells produced the most (UVB: 1,296 pg/ml). When co-exposed to IL1α, irradiated HaCaT had the greatest fold of TNFα release (UVB: 16.2-fold, UVA + B: 8.9-fold and UVB + A: 6.1-fold). The pattern of activation and TNFα secretion of HaCaT cells mirrored that of Colo16 cells. It is likely that the presence of molecular alterations in HaCaT cells may be responsible for its different responses to that seen for HEK cells. The results of this study suggest caution in using HaCaT cells as a substitute for normal keratinocytes in investigating UV-induced cells signalling pathways.
Tài liệu tham khảo
Arany I, Brysk MM, Chen Z, Adler-Storthz K, Tyring SK (1999) Differentiation-dependent expression of growth regulatory cytokines and their receptors in squamous cell carcinomas of the head and neck. Anticancer Res 19(4C):3623–3626
Bari L, Bacsa S, Sonkoly E, Bata-Csorgo Z, Kemeny L, Dobozy A, Szell M (2011) Comparison of stress-induced PRINS gene expression in normal human keratinocytes and HaCaT cells. Arch Dermatol Res 303(10):745–752
Bashir MM, Sharma MR, Werth VP (2009) TNF-α production in the skin. Arch Dermatol Res 301(1):87–91
Bashir MM, Sharma MR, Werth VP (2009) UVB and proinflammatory cytokines synergistically activate TNF-α production in keratinocytes through enhanced gene transcription. J Invest Dermatol 129(4):994–1001
Bode AM, Dong Z (2003) Mitogen-activated protein kinase activation in UV-induced signal transduction. Sci STKE (167):RE2
Boukamp P (2005) Non-melanoma skin cancer: what drives tumor development and progression? Carcinogenesis 26(10):1657–1667
Boukamp P (2005) UV-induced skin cancer: similarities–variations. J Dtsch Dermatol Ges 3(7):493–503
Boukamp P, Petrussevska RT, Breitkreutz D, Hornung J, Markham A, Fusenig NE (1988) Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol 106(3):761–771
Chaturvedi V, Qin JZ, Denning MF, Choubey D, Diaz MO, Nickoloff BJ (2001) Abnormal NF-κB signaling pathway with enhanced susceptibility to apoptosis in immortalized keratinocytes. J Dermatol Sci 26(1):67–78
Christmann M, Tomicic MT, Aasland D, Kaina B (2007) A role for UV-light-induced c-Fos: stimulation of nucleotide excision repair and protection against sustained JNK activation and apoptosis. Carcinogenesis 28(1):183–190
Clingen PH, Berneburg M, Petit-Frere C, Woollons A, Lowe JE, Arlett CF, Green MH (2001) Contrasting effects of an ultraviolet B and an ultraviolet A tanning lamp on interleukin-6, tumour necrosis factor-α and intercellular adhesion molecule-1 expression. Br J Dermatol 145(1):54–62
Clydesdale GJ, Dandie GW, Muller HK (2001) Ultraviolet light induced injury: immunological and inflammatory effects. Immunol Cell Biol 79(6):547–568
Dajee M, Lazarov M, Zhang JY, Cai T, Green CL, Russell AJ, Marinkovich MP, Tao S, Lin Q, Kubo Y, Khavari PA (2003) NF-κB blockade and oncogenic Ras trigger invasive human epidermal neoplasia. Nature 421(6923):639–643
Dazard JE, Gal H, Amariglio N, Rechavi G, Domany E, Givol D (2003) Genome-wide comparison of human keratinocyte and squamous cell carcinoma responses to UVB irradiation: implications for skin and epithelial cancer. Oncogene 22(19):2993–3006
Devalaraja MN, Wang DZ, Ballard DW, Richmond A (1999) Elevated constitutive IκB kinase activity and IκB-α phosphorylation in Hs294T melanoma cells lead to increased basal MGSA/GRO-α transcription. Cancer Res 59(6):1372–1377
Dicker AJ, Serewko MM, Dahler AL, Khanna KK, Kaur P, Li A, Strutton GM, Saunders NA (2000) Functional characterization of cultured cells derived from an intraepidermal carcinoma of the skin (IEC-1). Exp Cell Res 258(2):352–360
Duthie MS, Kimber I, Norval M (1999) The effects of ultraviolet radiation on the human immune system. Br J Dermatol 140(6):995–1009
Faurschou A, Gniadecki R, Calay D, Wulf HC (2008) TNF-α impairs the S-G2/M cell cycle checkpoint and cyclobutane pyrimidine dimer repair in premalignant skin cells: role of the PI3K-Akt pathway. J Invest Dermatol 128(8):2069–2077
Godar DE (2005) UV doses worldwide. Photochem Photobiol 81(4):736–749
Gross ND, Boyle JO, Du B, Kekatpure VD, Lantowski A, Thaler HT, Weksler BB, Subbaramaiah K, Dannenberg AJ (2007) Inhibition of Jun NH2-terminal kinases suppresses the growth of experimental head and neck squamous cell carcinoma. Clin Cancer Res 13(19):5910–5917
Halliday GM, Lyons JG (2008) Inflammatory doses of UV may not be necessary for skin carcinogenesis. Photochem Photobiol 84(2):272–283
Hazzalin CA, Le Panse R, Cano E, Mahadevan LC (1998) Anisomycin selectively desensitizes signalling components involved in stress kinase activation and fos and jun induction. Mol Cell Biol 18(4):1844–1854
He YY, Huang JL, Chignell CF (2004) Delayed and sustained activation of extracellular signal-regulated kinase in human keratinocytes by UVA: implications in carcinogenesis. J Biol Chem 279(51):53867–53874
Huynh TT, Chan KS, Piva TJ (2009) Effect of ultraviolet radiation on the expression of pp 38MAPK and furin in human keratinocyte-derived cell lines. Photodermatol Photoimmunol Photomed 25(1):20–29
Ishida T, Sakaguchi I (2007) Protection of human keratinocytes from UVB-induced inflammation using root extract of Lithospermum erythrorhizon. Biol Pharm Bull 30(5):928–934
Junttila MR, Ala-Aho R, Jokilehto T, Peltonen J, Kallajoki M, Grenman R, Jaakkola P, Westermarck J, Kahari VM (2007) p38α and p38δ mitogen-activated protein kinase isoforms regulate invasion and growth of head and neck squamous carcinoma cells. Oncogene 26(36):5267–5279
Krohne TU, Hunt S, Holz FG (2009) Effect of 308 nm excimer laser irradiation on retinal pigment epithelium cell viability in vitro. Br J Ophthalmol 93(1):91–95
Kuchel JM, Barnetson RS, Halliday GM (2002) Ultraviolet A augments solar-simulated ultraviolet radiation-induced local suppression of recall responses in humans. J Invest Dermatol 118(6):1032–1037
La E, Muga SJ, Locniskar MF, Fischer SM (1999) Altered expression of interleukin-1 receptor antagonist in different stages of mouse skin carcinogenesis. Mol Carcinog 24(4):276–286
Larsson P, Ollinger K, Rosdahl I (2006) Ultraviolet (UV)A- and UVB-induced redox alterations and activation of nuclear factor-κB in human melanocytes-protective effects of α-tocopherol. Br J Dermatol 155(2):292–300
Latonen L, Laiho M (2005) Cellular UV damage responses–functions of tumor suppressor p53. Biochim Biophys Acta 1755(2):71–89
Latonen L, Taya Y, Laiho M (2001) UV-radiation induces dose-dependent regulation of p53 response and modulates p53-HDM2 interaction in human fibroblasts. Oncogene 20(46):6784–6793
Lewis DA, Hengeltraub SF, Gao FC, Leivant MA, Spandau DF (2006) Aberrant NF-κB activity in HaCaT cells alters their response to UVB signaling. J Invest Dermatol 126(8):1885–1892
Lopez-Camarillo C, Ocampo EA, Casamichana ML, Perez-Plasencia C, Alvarez-Sanchez E, Marchat LA (2012) Protein kinases and transcription factors activation in response to UV-radiation of skin: implications for carcinogenesis. Int J Mol Sci 13(1):142–172
Marionnet AV, Chardonnet Y, Viac J, Schmitt D (1997) Differences in responses of interleukin-1 and tumor necrosis factor α production and secretion to cyclosporin-A and ultraviolet B-irradiation by normal and transformed keratinocyte cultures. Exp Dermatol 6(1):22–28
Micallef L, Belaubre F, Pinon A, Jayat-Vignoles C, Delage C, Charveron M, Simon A (2009) Effects of extracellular calcium on the growth-differentiation switch in immortalized keratinocyte HaCaT cells compared with normal human keratinocytes. Exp Dermatol 18(2):143–151
Mikita N, Kanazawa N, Yoshimasu T, Ikeda T, Li HJ, Yamamoto Y, Furukawa F (2009) The protective effects of ultraviolet A1 irradiation on spontaneous lupus erythematosus-like skin lesions in MRL/lpr mice. Clin Dev Immunol 2009:673952
Moise AF, Gies HP, Harrison SL (1999) Estimation of the annual solar UVR exposure dose of infants and small children in tropical Queensland, Australia. Photochem Photobiol 69(4):457–463
Muller C, Bektas M, Geilen CC (2003) Differential involvement of ceramide in TNFα-mediated activation of NF-κB in primary human keratinocytes and HaCaT keratinocytes. Cell Mol Biol 49:399–407
Muthusamy V, Hodges LD, Macrides TA, Boyle GM, Piva TJ (2011) Effect of novel marine nutraceuticals on IL-1α-mediated TNF-α release from UVB-irradiated human melanocyte-derived cells. Oxid Med Cell Longev 2011:728645
Muthusamy V, Piva TJ (2010) The UV response of the skin: a review of the MAPK, NFκB and TNFα signal transduction pathways. Arch Dermatol Res 302(1):5–17
Nghiem DX, Kazimi N, Clydesdale G, Ananthaswamy HN, Kripke ML, Ullrich SE (2001) Ultraviolet A radiation suppresses an established immune response: implications for sunscreen design. J Invest Dermatol 117(5):1193–1199
Olschlager V, Schrader A, Hockertz S (2009) Comparison of primary human fibroblasts and keratinocytes with immortalized cell lines regarding their sensitivity to sodium dodecyl sulfate in a neutral red uptake cytotoxicity assay. Arzneimittelforschung 59(3):146–152
Pastore S, Lulli D, Potapovich AI, Fidanza P, Kostyuk VA, Dellambra E, De Luca C, Maurelli R, Korkina LG (2011) Differential modulation of stress-inflammation responses by plant polyphenols in cultured normal human keratinocytes and immortalized HaCaT cells. J Dermatol Sci 63(2):104–114
Petit-Frere C, Capulas E, Lyon DA, Norbury CJ, Lowe JE, Clingen PH, Riballo E, Green MH, Arlett CF (2000) Apoptosis and cytokine release induced by ionizing or ultraviolet B radiation in primary and immortalized human keratinocytes. Carcinogenesis 21(6):1087–1095
Ravi D, Muniyappa H, Das KC (2008) Caffeine inhibits UV-mediated NF-κB activation in A2058 melanoma cells: an ATM-PKCδ-p38 MAPK-dependent mechanism. Mol Cell Biochem 308(1–2):193–200
Schieke SM, Ruwiedel K, Gers-Barlag H, Grether-Beck S, Krutmann J (2005) Molecular crosstalk of the ultraviolet A and ultraviolet B signaling responses at the level of mitogen-activated protein kinases. J Invest Dermatol 124(4):857–859
Schneider G, Kramer OH (2011) NFκB/p53 crosstalk-a promising new therapeutic target. Biochim Biophys Acta 1815(1):90–103
Schoop VM, Mirancea N, Fusenig NE (1999) Epidermal organization and differentiation of HaCaT keratinocytes in organotypic coculture with human dermal fibroblasts. J Invest Dermatol 112(3):343–353
Schurer N, Kohne A, Schliep V, Barlag K, Goerz G (1993) Lipid composition and synthesis of HaCaT cells, an immortalized human keratinocyte line, in comparison with normal human adult keratinocytes. Exp Dermatol 2(4):179–185
Soufir N, Moles JP, Vilmer C, Moch C, Verola O, Rivet J, Tesniere A, Dubertret L, Basset-Seguin N (1999) P16 UV mutations in human skin epithelial tumors. Oncogene 18(39):5477–5481
Squarize CH, Castilho RM, Sriuranpong V, Pinto DS Jr, Gutkind JS (2006) Molecular cross-talk between the NFκB and STAT3 signaling pathways in head and neck squamous cell carcinoma. Neoplasia 8(9):733–746
Starcher B (2000) Role for tumour necrosis factor-α receptors in ultraviolet-induced skin tumours. Br J Dermatol 142(6):1140–1147
Van Laethem A, Nys K, Van Kelst S, Claerhout S, Ichijo H, Vandenheede JR, Garmyn M, Agostinis P (2006) Apoptosis signal regulating kinase-1 connects reactive oxygen species to p38 MAPK-induced mitochondrial apoptosis in UVB-irradiated human keratinocytes. Free Radic Biol Med 41(9):1361–1371
Wang XY, Bi ZG (2006) UVB-irradiated human keratinocytes and interleukin-1α indirectly increase MAP kinase/AP-1 activation and MMP-1 production in UVA-irradiated dermal fibroblasts. Chin Med J 119(10):827–831
Werth VP, Zhang W (1999) Wavelength-specific synergy between ultraviolet radiation and interleukin-1 α in the regulation of matrix-related genes: mechanistic role for tumor necrosis factor-α. J Invest Dermatol 113(2):196–201
Whiteside JR, McMillan TJ (2009) A bystander effect is induced in human cells treated with UVA radiation but not UVB radiation. Radiat Res 171(2):204–211