Cargando…
Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis
BACKGROUND: Metastatic malignant melanoma is one of the most aggressive malignancies and its treatment remains challenging. Recent studies demonstrate that the melanoma metastasis has correlations with the heightened activations of protein kinase C ζ (PKCζ) and cyclooxygenase-2 (COX-2) signaling pat...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5581453/ https://www.ncbi.nlm.nih.gov/pubmed/28865485 http://dx.doi.org/10.1186/s13046-017-0585-2 |
_version_ | 1783261051871035392 |
---|---|
author | Zhou, Ping Qin, Jiaqi Li, Yuan Li, Guoxia Wang, Yinsong Zhang, Ning Chen, Peng Li, Chunyu |
author_facet | Zhou, Ping Qin, Jiaqi Li, Yuan Li, Guoxia Wang, Yinsong Zhang, Ning Chen, Peng Li, Chunyu |
author_sort | Zhou, Ping |
collection | PubMed |
description | BACKGROUND: Metastatic malignant melanoma is one of the most aggressive malignancies and its treatment remains challenging. Recent studies demonstrate that the melanoma metastasis has correlations with the heightened activations of protein kinase C ζ (PKCζ) and cyclooxygenase-2 (COX-2) signaling pathways. Targeted inhibitions for PKCζ and COX-2 have been considered as the promising strategies for the treatment of melanoma metastasis. Thus, the PKCζ inhibitor J-4 and COX-2 inhibitor Celecoxib were combined to treat melanoma metastasis in this study. METHODS: The Transwell assay, Wound-healing assay and Adhesion assay were used to evaluate the inhibition of combined therapy of J-4 and Celecoxib on melanoma cells invasion, migration and adhesion in vitro, respectively. The impaired actin polymerization was observed by confocal microscope and inactivated signal pathways about PKCζ and COX-2 were confirmed by the Western blotting assay. The B16-F10/C57BL mouse melanoma model was used to test the inhibition of combined therapy of J-4 and Celecoxib on melanoma metastasis in vivo. RESULTS: The in vitro results showed that the combination of J-4 and Celecoxib exerted synergistic inhibitory effects on the migration, invasion and adhesion of melanoma B16-F10 and A375 cells with combination index less than 1. The actin polymerization and phosphorylation of Cofilin required in cell migration were severely impaired, which is due to the inactivation of PKCζ related signal pathways and the decrease of COX-2. The combined inhibition of PKCζ and COX-2 induced Mesenchymal-Epithelial Transition (MET) in melanoma cells with the expression of E-Cadherin increasing and Vimentin decreasing. The secretion of MMP-2/MMP-9 also significantly decreased after the combination treatment. In C57BL/6 mice intravenously injected with B16-F10 cells (5 × 10(4) cells/mouse), co-treatment of J-4 and Celecoxib also severely suppressed melanoma lung metastasis. The body weight monitoring and HE staining results indicated the low toxicity of the combination therapy. CONCLUSIONS: This study demonstrates that the combination therapy of PKCζ and COX-2 inhibitors can significantly inhibit melanoma metastasis in vitro and in vivo, which will be an efficient strategy for treatment of melanoma metastasis in clinics. |
format | Online Article Text |
id | pubmed-5581453 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-55814532017-09-06 Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis Zhou, Ping Qin, Jiaqi Li, Yuan Li, Guoxia Wang, Yinsong Zhang, Ning Chen, Peng Li, Chunyu J Exp Clin Cancer Res Research BACKGROUND: Metastatic malignant melanoma is one of the most aggressive malignancies and its treatment remains challenging. Recent studies demonstrate that the melanoma metastasis has correlations with the heightened activations of protein kinase C ζ (PKCζ) and cyclooxygenase-2 (COX-2) signaling pathways. Targeted inhibitions for PKCζ and COX-2 have been considered as the promising strategies for the treatment of melanoma metastasis. Thus, the PKCζ inhibitor J-4 and COX-2 inhibitor Celecoxib were combined to treat melanoma metastasis in this study. METHODS: The Transwell assay, Wound-healing assay and Adhesion assay were used to evaluate the inhibition of combined therapy of J-4 and Celecoxib on melanoma cells invasion, migration and adhesion in vitro, respectively. The impaired actin polymerization was observed by confocal microscope and inactivated signal pathways about PKCζ and COX-2 were confirmed by the Western blotting assay. The B16-F10/C57BL mouse melanoma model was used to test the inhibition of combined therapy of J-4 and Celecoxib on melanoma metastasis in vivo. RESULTS: The in vitro results showed that the combination of J-4 and Celecoxib exerted synergistic inhibitory effects on the migration, invasion and adhesion of melanoma B16-F10 and A375 cells with combination index less than 1. The actin polymerization and phosphorylation of Cofilin required in cell migration were severely impaired, which is due to the inactivation of PKCζ related signal pathways and the decrease of COX-2. The combined inhibition of PKCζ and COX-2 induced Mesenchymal-Epithelial Transition (MET) in melanoma cells with the expression of E-Cadherin increasing and Vimentin decreasing. The secretion of MMP-2/MMP-9 also significantly decreased after the combination treatment. In C57BL/6 mice intravenously injected with B16-F10 cells (5 × 10(4) cells/mouse), co-treatment of J-4 and Celecoxib also severely suppressed melanoma lung metastasis. The body weight monitoring and HE staining results indicated the low toxicity of the combination therapy. CONCLUSIONS: This study demonstrates that the combination therapy of PKCζ and COX-2 inhibitors can significantly inhibit melanoma metastasis in vitro and in vivo, which will be an efficient strategy for treatment of melanoma metastasis in clinics. BioMed Central 2017-09-02 /pmc/articles/PMC5581453/ /pubmed/28865485 http://dx.doi.org/10.1186/s13046-017-0585-2 Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Zhou, Ping Qin, Jiaqi Li, Yuan Li, Guoxia Wang, Yinsong Zhang, Ning Chen, Peng Li, Chunyu Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis |
title | Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis |
title_full | Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis |
title_fullStr | Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis |
title_full_unstemmed | Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis |
title_short | Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis |
title_sort | combination therapy of pkcζ and cox-2 inhibitors synergistically suppress melanoma metastasis |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5581453/ https://www.ncbi.nlm.nih.gov/pubmed/28865485 http://dx.doi.org/10.1186/s13046-017-0585-2 |
work_keys_str_mv | AT zhouping combinationtherapyofpkczandcox2inhibitorssynergisticallysuppressmelanomametastasis AT qinjiaqi combinationtherapyofpkczandcox2inhibitorssynergisticallysuppressmelanomametastasis AT liyuan combinationtherapyofpkczandcox2inhibitorssynergisticallysuppressmelanomametastasis AT liguoxia combinationtherapyofpkczandcox2inhibitorssynergisticallysuppressmelanomametastasis AT wangyinsong combinationtherapyofpkczandcox2inhibitorssynergisticallysuppressmelanomametastasis AT zhangning combinationtherapyofpkczandcox2inhibitorssynergisticallysuppressmelanomametastasis AT chenpeng combinationtherapyofpkczandcox2inhibitorssynergisticallysuppressmelanomametastasis AT lichunyu combinationtherapyofpkczandcox2inhibitorssynergisticallysuppressmelanomametastasis |