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Premature keratinocyte death and expression of marker proteins of apoptosis in human skin after UVB exposure

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Abstract

Epidermal keratinocytes undergo a process of terminal differentiation or cornification that in many aspects resembles apoptosis. It is characterized by the elimination of cell nuclei within the granular layer, whereas the cytoplasm is transformed into horn cells. Premature death of keratinocytes can be induced by extrinsic factors such as UV irradiation. We investigated the time-dependent expression of apoptotic marker proteins in the skin of one healthy human volunteer after irradiation with a fourfold minimal erythema dose (MED) of UVB. The data were supplemented by including healthy skin areas of biopsies from patients UVB-irradiated for therapeutic reasons. Punch biopsies were analysed by in situ end-labelling (ISEL) for DNA strand breaks and by immunohistochemistry for expression of p53, bcl-2, active caspase-3 and its proform, and deoxyribonuclease I (DNase I). Keratinocytes with pyknotic nuclei were first detected 6 h after UVB exposure, and apoptotic keratinocytes (sunburn cells) 12 h after exposure. These aggregated to sunburn bodies after 24 h. In control skin, nuclei with DNA strand breaks were only occasionally detected in the granular layer but 6 h after UVB irradiation in the spinous layer. After 12 h, many sunburn cells were ISEL-positive and positively stained for active caspase-3, P53, and DNase I. Morphometric evaluation of the immunohistochemical data demonstrated that maximal upregulation of P53, DNase I and activation of caspase-3 occurred 12 h after irradiation and in advance of the peak of apoptotic cell death reached after 24 h as verified by ISEL. In contrast, strong Bcl-2 immunostaining appeared restricted to presumed melanocytes and basal cells but was not increased after UVB irradiation.

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References

  1. Niemann C, Watt FM (2002) Designer skin: lineage commitment in postnatal epidermis. Trends Cell Biol 12:185–192

    Article  CAS  PubMed  Google Scholar 

  2. Candi E, Oddi S, Terrinoni A, Paradisi A, Ranalli M, Finazzi-Agró A, Melino G (2001) Transglutaminase 5 cross-links loricrin, involucrin, and small proline-rich proteins in vitro. J Biol Chem 276:35014–35023

    Article  CAS  PubMed  Google Scholar 

  3. Fesus L, Davies PJA, Piacentini M (1991) Apoptosis: molecular mechanisms in programmed cell death. Eur J Cell Biol 56:170–177

    CAS  PubMed  Google Scholar 

  4. Polzar B, Zanotti S, Stephan H, Rauch F, Peitsch MC, Irmler M, Tschopp J, Mannherz HG (1994) The correlation of the distribution of deoxyribonuclease I in rat tissues and its correlation to cellular turnover and apoptosis (programmed cell death). Eur J Cell Biol 64:200–210

    Google Scholar 

  5. Lippens S, Kockx M, Knaapen M, Mortier L, Polakowska R, Verheyen A, Garmyn M, Zwijsen A, Formstecher P, Huylebroeck D, Vandenabeele P, Declercq W (2000) Epidermal differentiation does not involve the pro-apoptotic executioner caspases, but is associated with caspase-14 induction and processing. Cell Death Differ 7:1218–1224

    Article  CAS  PubMed  Google Scholar 

  6. Matsui MS, DeLeo VA (1991) Longwave ultraviolet radiation and promotion of skin cancer. Cancer Cells 3:8–12

    CAS  PubMed  Google Scholar 

  7. Tornaletti S, Pfeiffer GP (1996) UV damage and repair mechanisms in mammalian cells. Bioessays 18:221–228

    CAS  PubMed  Google Scholar 

  8. Brash DE (1997) Sunlight and the onset of skin cancer. Trends Genet 13:410–414

    Article  CAS  PubMed  Google Scholar 

  9. Berner F, Asselineau D (1998) UVA exposure of human skin reconstituted in vitro induces apoptosis of dermal fibroblasts: subsequent connective tissue repair and implications in photoaging. Cell Death Differ 5:792–802

    Article  PubMed  Google Scholar 

  10. von Kobyletzki G, Heine O, Stephan H, Pieck C, Stücker M, Hoffmann K, Altmeyer P, Mannherz HG (2000) UVA1 irradiation induces apoptosis in cutaneous T-cell lymphoma in vivo. Photodermatol Photoimmunol Photomed 16:271–277

    PubMed  Google Scholar 

  11. Liu M, Pelling JC (1995) UV-B/A irradiation of mouse keratinocytes results in p53-mediated WAF1/cip1 expression. Oncogene 10:1955–1960

    CAS  PubMed  Google Scholar 

  12. Kulms D, Schwarz T (2000) Molecular mechanisms of UV-induced apoptosis. Photodermatol Photoimmunol Photomed 16:195–201

    Article  CAS  PubMed  Google Scholar 

  13. Murphy G, Young AR, Wulf HC, Kulms D, Schwarz T (2001) The molecular determinants of sunburn cell formation. Exp Dermatol 10:155–160

    Article  CAS  PubMed  Google Scholar 

  14. Rehemtulla A, Hamilton CA, Chinnaiyan AM, Dixit VM (1997) Ultraviolet radiation-induced apoptosis is mediated by activation of CD95 (Fas/APO-1). J Biol Chem 272:25783–25786

    Article  CAS  PubMed  Google Scholar 

  15. Aragane Y, Kulms D, Metze D, Wilkes G, Pöppelmann B, Luger TA, Schwarz T (1998) Ultraviolet light induces apoptosis via direct activation of CD95 (Fas/APO-1) independently of its ligand CD95L. J Cell Biol 140:171–182

    Article  CAS  PubMed  Google Scholar 

  16. Filipowicz E, Adegboyega P, Sanchez RL, Gatalica Z (2002) Expression of CD95 (Fas) in sun-exposed human skin and cutaneous carcinomas. Cancer 94:814–819

    Article  CAS  PubMed  Google Scholar 

  17. Haimovitz-Friedman A, Kann C-C, Ehleiter D, Persaud RS, McLoughlin M, Fuks Z, Kolesnick RN (1994) Ionizing radiation acts on cellular membranes to generate ceramide and initiate apoptosis. J Exp Med 180:525–535

    CAS  PubMed  Google Scholar 

  18. Iwasaki K, Izawa M, Mihara M (1996) UV-induced apoptosis in rat skin. J Dermatol Sci 12:31–35

    Article  CAS  PubMed  Google Scholar 

  19. Sheehan JM, Young AR (2000) Ultraviolet radiation induced apoptosis in human skin in vivo. Radiat Protect Dosim 91:47–50

    CAS  Google Scholar 

  20. Young AR (1986) The sunburn cell. Photodermatology 4:127–134

    Google Scholar 

  21. Taylor JK, Quing ZQ, Monia BP, Marcusson EG, Dean NM (1999) Inhibition of Bcl-XL expression sensitizes normal human keratinocytes and epithelial cells to apoptotic stimuli. Oncogene 18:4495–4504

    Article  CAS  PubMed  Google Scholar 

  22. Schendel SL, Montal M, Reed JC (1998) Bcl-2 family proteins as ion-channels. Cell Death Differ 5:372–380

    CAS  PubMed  Google Scholar 

  23. Earnshaw WC, Martins LM, Kaufmann SH (1999) Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu Rev Biochem 68:383–424

    CAS  PubMed  Google Scholar 

  24. Lamkanfi M, Declercq W, Kalai M, Saelens X, Vandenabeele P (2002) Alice in caspase land. A phylogenetic analysis of caspases from worm to man. Cell Death Differ 9:358–361

    Article  CAS  PubMed  Google Scholar 

  25. Wyllie AH (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284:555–556

    CAS  PubMed  Google Scholar 

  26. Peitsch MC, Mannherz HG, Tschopp J (1994) The apoptosis endonucleases: cleaning up after cell death. Trends Cell Biol 4:37–41

    Article  CAS  Google Scholar 

  27. McIllroy D, Tanaka M, Sakahira H, Fukuyama H, Suzuki M, Yamamura K, Ohsawa Y, Uchiyama Y, Nagata S (2000) An auxiliary mode of apoptotic DNA fragmentation provided by phagocytes. Genes Dev 14:549–558

    PubMed  Google Scholar 

  28. Enari M, Sakahira H, Yoloyama H, Okawa K, Iwamatsu A, Nagata S (1997) A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391:43–50

    Google Scholar 

  29. Mukae K, Enari M, Sakahira H, Fukuda Y, Inazawa J, Toh H, Nagata S (1998) Molecular cloning and characterization of human caspase-activated DNase. Proc Natl Acad Sci U S A 95:9123–9128

    Article  CAS  PubMed  Google Scholar 

  30. Peitsch MC, Polzar B, Stephan H, Crompton T, McDonald HR, Mannherz HG, Tschopp J (1993) Characterization of the endogenous deoxyribonuclease involved in nuclear DNA degradation during apoptosis (programmed cell death). EMBO J 12:371–377

    CAS  PubMed  Google Scholar 

  31. Napirei M, Karsunky H, Zevnik B, Stephan H, Mannherz HG, Möröy T (2000) Features of systemic Lupus erythematosus (SLE) in Dnase 1 deficient mice. Nat Genet 25:177–181

    Article  CAS  PubMed  Google Scholar 

  32. Shiokawa D, Ohyama H, Yamada T, Takahashi K, Tanuma S (1994) Identification of an endonuclease responsible for apoptosis in rat thymocytes. Eur J Biochem 226:23–30

    CAS  PubMed  Google Scholar 

  33. Liu QY, Pandey S, Singh RK, Lin W, Ribecco M, Borowy-Borowski H, Smith B, LeBlanc J, Walker R, Sikorska M (1998) DNaseY: a rat DNase I-like gene coding for a constitutively expressed chromatin-bound endonuclease. Biochemistry 37:10134–10143

    Article  CAS  PubMed  Google Scholar 

  34. Shiokawa D, Tanuma S (1998) Molecular cloning and expression of a cDNA encoding an apoptotic endonuclease DNase γ. Biochem J 332:713–720

    CAS  PubMed  Google Scholar 

  35. Fitzpatrick TB (1988) The validity and practicality of sun-reactive skin types I and IV. Arch Dermatol 124:869–887

    Article  CAS  PubMed  Google Scholar 

  36. Rauch F, Polzar B, Stephan H, Zanotti S, Paddenberg R, Mannherz HG (1997) Androgen ablation leads to an upregulation and intranuclear accumulation of deoxyribonuclease I in rat prostate epithelial cells paralleling their apoptotic elimination. J Cell Biol 137:909–923

    Article  CAS  PubMed  Google Scholar 

  37. Gavrieli Y, Sherman Y, Ben-Sasson SA (1992) Identification of programmed cell death in situ via specific labelling of nuclear DNA fragmentation. J Cell Biol 119:493–501

    CAS  PubMed  Google Scholar 

  38. Wijsman JH, Jonker RR, Keijzer R, van de Velde CJH, Cornelisse CJ, van Dierendonck JH (1993) A new method to detect apoptosis in paraffin sections: in situ end-labelling of fragmented DNA. J Histochem Cytochem 41:7–12

    CAS  PubMed  Google Scholar 

  39. Polakowska RR, Piacentini M, Bartelett R, Goldsmith LA, Haake AR (1994) Apoptosis in human skin development: morphogenesis, periderm, and stem cells. Dev Dyn 199:176–188

    CAS  Google Scholar 

  40. Breuckmann F, Pieck, C, Kreuter A, Bacharach-Buhles M, Mannherz HG, Altmeyer P, von Kobyletzki G (2001) Opposing effects of UVA1 phototherapy on the expression of bcl-2 and p53 in atopic dermatitis. Arch Dermatol Res 293:178–183

    CAS  PubMed  Google Scholar 

  41. Zanotti S, Fisseler-Eckhoff A, Mannherz HG (2003) Changes in the topological expression of markers of differentiation and apoptosis in defined stages of human cervical dysplasia and carcinoma. Gynecol Oncol (in press)

  42. Ponten F, Berne B, Ren ZP, Nister M, Ponten J (1995) Ultraviolet light induces expression of p53 and p21 in human skin: effect of sunscreen and constitutive p21 expression in skin appendages. J Invest Dermatol 105:402–406

    CAS  PubMed  Google Scholar 

  43. Qin, J-Z, Chaturvedi V, Denning M, Bacon P, Panella J, Choubey D, Nickoloff BJ (2002) Regulation of apoptosis by p53 in UV-irradiated human epidermis, psoriatic plaques and senescent keratinocytes. Oncogene 21:2991–3002

    Article  CAS  PubMed  Google Scholar 

  44. Bohlinger I, Leist M, Gantner F, Angermüller S, Tiegs G, Wendel A (1996) DNA fragmentation in mouse organs during endotoxic shock. Am J Pathol 149:1381–1393

    CAS  PubMed  Google Scholar 

  45. Bowen AR, Hanks AN, Allen SM, Alexander A, Diedrich MJ, Grossman D (2003) Apoptosis regulators and responses in human melanocytic and keratinocytic cells. J Invest Dermatol 120:48–55

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We gratefully acknowledge the financial support of the DFG (Deutsche Forschungsgemeinschaft, Bonn, Germany) and of the Medical Faculty of the Ruhr-University Bochum via a grant from FoRUM. This work is part of the doctoral thesis of P. Maß.

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Authors and Affiliations

  1. Department of Anatomy and Embryology, Ruhr-University Bochum, Universitätsstr. 150, 44780, Bochum, Germany

    Peter Maß & Hans Georg Mannherz

  2. Department of Dermatology, St. Josef Hospital, Ruhr-University, Bochum, Germany

    Klaus Hoffmann, Thilo Gambichler & Peter Altmeyer

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  1. Peter Maß
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Correspondence to Hans Georg Mannherz.

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Maß, P., Hoffmann, K., Gambichler, T. et al. Premature keratinocyte death and expression of marker proteins of apoptosis in human skin after UVB exposure. Arch Dermatol Res 295, 71–79 (2003). https://doi.org/10.1007/s00403-003-0403-x

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