17-Aminogeldanamycin Inhibits Constitutive Nuclear Factor-Kappa B (NF-κB) Activity in Patient-Derived Melanoma Cell Lines
| Autor: | Magdalena Rogut, Malgorzata Czyz, Mariusz L. Hartman, Michal Wozniak, Aleksandra Mielczarek-Lewandowska |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Vascular Endothelial Growth Factor A
HSP90 inhibitor Lactams Macrocyclic cyclin D1 NRAS Catalysis Article Flow cytometry BRAF Inorganic Chemistry lcsh:Chemistry chemistry.chemical_compound Cyclin D1 Heat shock protein BCL-XL Cell Line Tumor medicine Benzoquinones Tumor Cells Cultured melanoma Humans p65/NF-κB Interleukin 8 Physical and Theoretical Chemistry CHUK Molecular Biology lcsh:QH301-705.5 Spectroscopy medicine.diagnostic_test IL-8 Melanoma Organic Chemistry Interleukin-8 NF-kappa B General Medicine medicine.disease 17-aminogeldanamycin Molecular biology NFKBIE VEGF Computer Science Applications Vascular endothelial growth factor Gene Expression Regulation Neoplastic Toll-Like Receptor 4 chemistry lcsh:Biology (General) lcsh:QD1-999 |
| Zdroj: | International Journal of Molecular Sciences Volume 21 Issue 11 International Journal of Molecular Sciences, Vol 21, Iss 3749, p 3749 (2020) |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms21113749 |
| Popis: | Melanoma remains incurable skin cancer, and targeting heat shock protein 90 (HSP90) is a promising therapeutic approach. In this study, we investigate the effect of 17-aminogeldanamycin, a potent HSP90 inhibitor, on nuclear factor-kappa B (NF-&kappa B) activity in BRAFV600E and NRASQ61R patient-derived melanoma cell lines. We performed time-lapse microscopy and flow cytometry to monitor changes in cell confluence and viability. The NF-&kappa B activity was determined by immunodetection of phospho-p65 and assessment of expression of NF-&kappa B-dependent genes by quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, and enzyme-linked immunosorbent assay (ELISA). Constitutive activity of p65/NF-&kappa B was evident in all melanoma cell lines. Differences in its level might be associated with genetic alterations in CHUK, IL1B, MAP3K14, NFKBIE, RIPK1, and TLR4, while differences in transcript levels of NF-&kappa B-inducible genes revealed by PCR array might result from the contribution of other regulatory mechanisms. 17-Aminogeldanamycin markedly diminished the level of phospho-p65, but the total p65 protein level was unaltered, indicating that 17-aminogeldanamycin inhibited activation of p65/NF-&kappa B. This conclusion was supported by significantly reduced expression of selected NF-&kappa B-dependent genes: cyclin D1 (CCND1), C-X-C motif chemokine ligand 8 (CXCL8), and vascular endothelial growth factor (VEGF), as shown at transcript and protein levels, as well as secretion of IL-8 and VEGF. Our study indicates that 17-aminogeldanamycin can be used for efficient inhibition of NF-&kappa B activity and the simultaneous diminution of IL-8 and VEGF levels in the extracellular milieu of melanoma. |
| Databáze: | OpenAIRE |
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