Electrolytically generated acid functional water inhibits NF-κB activity by attenuating nuclear-cytoplasmic shuttling of p65 and p50 subunits
| Autor: | Masatake Asano, Masafumi Kusunoki, Yukina Nishio, Hirotaka Ota, Takahiro Gojoubori, Satoko Iwasa, Yasuhide Kaneko, Kensuke Nishio |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
P50 Transcription Genetic Biology Biochemistry Electrolysis 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cell Line Tumor Gene expression Humans Luciferase Secretion Interleukin 8 Molecular Biology Cell Nucleus 030102 biochemistry & molecular biology Interleukin-8 NF-kappa B Transcription Factor RelA Water NF-κB Cell Biology Molecular biology Blot Protein Subunits Protein Transport chemistry Cell culture 030220 oncology & carcinogenesis |
| Zdroj: | Journal of Receptors and Signal Transduction. 36:248-253 |
| ISSN: | 1532-4281 1079-9893 |
| DOI: | 10.3109/10799893.2015.1086883 |
| Popis: | Human β-defensin 2 (hBD2) gene expression is dependent on nuclear factor kappa B (NF-κB) activity. We have previously demonstrated that electrolytically generated acid functional water (FW) induces the expression of hBD2 in the human oral squamous cell carcinoma (OSCC) cell line Ca9-22. However, the induction was not dependent on NF-κB activity; in fact, FW inhibited NF-κB activity. Therefore, we hypothesized that FW might reduce spontaneous interleukin 8 (IL-8) secretion by Ca9-22 cells, which is heavily dependent on NF-κB activity. This study aimed at demonstrating the inhibitory effect of FW on NF-κB activity.Ca9-22 cells were incubated with FW, and spontaneous IL-8 secretion was observed by enzyme-linked immunosorbent assay. Luciferase assay was performed using the 5'-untranslated region of the IL-8 gene. The steps of NF-κB activation blocked by FW were evaluated by localization of the NF-κB subunits p65 and p50 by immunofluorescence staining. Western blotting was further performed to confirm the changes in NF-κB subunit localization.The Ca9-22 cells spontaneously secreted IL-8, which was rapidly and drastically inhibited by FW treatment. The luciferase assay demonstrated the inhibitory action of FW, which was diminished by deletion of the NF-κB binding site from this construct. FW treatment altered the distribution of both the p65 and p50 subunits. P65, which was localized in the nucleus during the resting state, moved to the cytoplasm after FW treatment, whereas, p50, localized in the cytoplasm during the resting state, moved to the nucleus subsequent to FW treatment.The results from this study indicate that FW might inhibit spontaneous IL-8 secretion by redistribution of the NF-κB subunits within the cells. |
| Databáze: | OpenAIRE |
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