Inverse Regulation of the Nuclear Factor-κB Binding to the p53 and Interleukin-8 κB Response Elements in Lesional Psoriatic Skin
| Autor: | Claus Johansen, Lars Iversen, Majken Westergaard, Karsten Kristiansen, Knud Kragballe, Jeanette Henningsen, Esben N. Flindt |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
p53
Adult Keratinocytes P50 Response element Blotting Western Fluorescent Antibody Technique Electrophoretic Mobility Shift Assay Dermatology Biology Protein Serine-Threonine Kinases Administration Cutaneous Response Elements Biochemistry NF-κB chemistry.chemical_compound Calcitriol Psoriasis medicine Humans Protein Isoforms Electrophoretic mobility shift assay RNA Messenger Transcription factor Molecular Biology Cells Cultured Skin IL-8 Kinase Reverse Transcriptase Polymerase Chain Reaction Interleukin-8 NF-kappa B DNA Cell Biology medicine.disease Molecular biology I-kappa B Kinase IκBα chemistry Gene Expression Regulation Immunology I-kappa B Proteins Dermatologic Agents Tumor Suppressor Protein p53 |
| Zdroj: | Journal of Investigative Dermatology. 124(6):1284-1292 |
| ISSN: | 0022-202X |
| DOI: | 10.1111/j.0022-202x.2005.23749.x |
| Popis: | Nuclear factor-kappaB (NF-kappaB) is an inducible nuclear transcription factor regulating a range of cellular processes. An imbalance of the DNA binding activity of NF-kappaB may, therefore, be part of the pathophysiological mechanisms in psoriasis. The purpose of this study was to determine the NF-kappaB DNA binding activity in psoriatic skin using three different kappaB sites and to determine how DNA binding activity was modulated by the anti-psoriatic drug calcipotriol. By electrophoretic mobility shift assay, we demonstrated that the NF-kappaB DNA binding to the p53 kappaB site was decreased, whereas the NF-kappaB DNA binding to the interleukin-8 (IL-8) kappaB site was increased in lesional psoriatic skin compared with non-lesional psoriatic skin. No regulation was seen on the NF-kappaB DNA binding to the major histocompatibility complex class I kappaB site. These changes were paralleled by a similar decrease in p53 expression and an increase in IL-8 expression in involved psoriatic skin compared with uninvolved skin as determined by quantitative RT-PCR. The alteration in NF-kappaB DNA binding activity was neither accompanied by any change in the expression of the inhibitor kappaB (IkappaB) kinases, IKKalpha, IKKbeta, and IKKgamma nor in the expression of the NF-kappaB inhibitor proteins, IkappaBalpha and IkappaBbeta. Immunofluorescence analysis revealed that p65 was sequestered in the cytoplasm of keratinocytes, whereas p50 exhibited a cytoplasmic as well as a nuclear localization. Interestingly, this distribution of p50 and p65 was similar in lesional and non-lesional psoriatic skin. Topical application of calcipotriol to lesional psoriatic skin for 4 d resulted in increased NF-kappaB binding to the p53 kappaB site and decreased NF-kappaB binding to the IL-8 kappaB site. Taken together, our data demonstrate that the NF-kappaB DNA binding activity is regulated in a specific manner in psoriatic skin depending on the kappaB sites investigated, and that topical treatment of psoriatic skin normalizes the abnormal NF-kappaB binding activity seen in lesional psoriatic skin. |
| Databáze: | OpenAIRE |
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