Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma
Abstract Polo-like kinase 1 (PLK1), a serine/threonine kinase, is overexpressed in melanoma and its expression has been associated with poor disease prognosis. PLK1 has been shown to interact with NUMB, a NOTCH antagonist. However, the exact role of PLK1, NUMB, and NOTCH signaling in epithelial-mese...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-01-01
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| Series: | npj Precision Oncology |
| Online Access: | https://doi.org/10.1038/s41698-023-00493-7 |
| _version_ | 1797363754737860608 |
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| author | Gagan Chhabra Chandra K. Singh Mary A. Ndiaye Shengqin Su Carl A. Shirley Nihal Ahmad |
| author_facet | Gagan Chhabra Chandra K. Singh Mary A. Ndiaye Shengqin Su Carl A. Shirley Nihal Ahmad |
| author_sort | Gagan Chhabra |
| collection | DOAJ |
| description | Abstract Polo-like kinase 1 (PLK1), a serine/threonine kinase, is overexpressed in melanoma and its expression has been associated with poor disease prognosis. PLK1 has been shown to interact with NUMB, a NOTCH antagonist. However, the exact role of PLK1, NUMB, and NOTCH signaling in epithelial-mesenchymal transition (EMT) in melanoma progression is unclear. In this study, Affymetrix microarray analysis was performed to determine differentially expressed genes following shRNA-mediated knockdown of PLK1 in human melanoma cells that showed significant modulations in EMT and metastasis-related genes. Using multiple PLK1-modulated melanoma cell lines, we found that PLK1 is involved in the regulation of cell migration, invasion, and EMT via its kinase activity and NOTCH activation. In vitro kinase assay and mass spectrometry analysis demonstrated a previously unknown PLK1 phosphorylation site (Ser413) on NUMB. Overexpression of non-phosphorylatable (S413A) and phosphomimetic (S413D) mutants of NUMB in melanoma cells implicated the involvement of NUMB-S413 phosphorylation in cell migration and invasion, which was independent of NOTCH activation. To determine the clinical relevance of these findings, immunohistochemistry was performed using melanoma tissue microarray, which indicated a strong positive correlation between PLK1 and N-cadherin, a protein required for successful EMT. These findings were supported by TCGA analysis, where expression of high PLK1 with low NUMB or high NOTCH or N-cadherin showed a significant decrease in survival of melanoma patients. Overall, these results suggest a potential role of PLK1 in EMT, migration, and invasion of melanoma cells. Our findings support the therapeutic targeting of PLK1, NUMB, and NOTCH for melanoma management. |
| first_indexed | 2024-03-08T16:24:33Z |
| format | Article |
| id | doaj.art-b16b349e73b64d49b30f12759328ed9c |
| institution | Directory Open Access Journal |
| issn | 2397-768X |
| language | English |
| last_indexed | 2024-03-08T16:24:33Z |
| publishDate | 2024-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | npj Precision Oncology |
| spelling | doaj.art-b16b349e73b64d49b30f12759328ed9c2024-01-07T12:06:41ZengNature Portfolionpj Precision Oncology2397-768X2024-01-018111510.1038/s41698-023-00493-7Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanomaGagan Chhabra0Chandra K. Singh1Mary A. Ndiaye2Shengqin Su3Carl A. Shirley4Nihal Ahmad5Department of Dermatology, University of WisconsinDepartment of Dermatology, University of WisconsinDepartment of Dermatology, University of WisconsinDepartment of Dermatology, University of WisconsinDepartment of Dermatology, University of WisconsinDepartment of Dermatology, University of WisconsinAbstract Polo-like kinase 1 (PLK1), a serine/threonine kinase, is overexpressed in melanoma and its expression has been associated with poor disease prognosis. PLK1 has been shown to interact with NUMB, a NOTCH antagonist. However, the exact role of PLK1, NUMB, and NOTCH signaling in epithelial-mesenchymal transition (EMT) in melanoma progression is unclear. In this study, Affymetrix microarray analysis was performed to determine differentially expressed genes following shRNA-mediated knockdown of PLK1 in human melanoma cells that showed significant modulations in EMT and metastasis-related genes. Using multiple PLK1-modulated melanoma cell lines, we found that PLK1 is involved in the regulation of cell migration, invasion, and EMT via its kinase activity and NOTCH activation. In vitro kinase assay and mass spectrometry analysis demonstrated a previously unknown PLK1 phosphorylation site (Ser413) on NUMB. Overexpression of non-phosphorylatable (S413A) and phosphomimetic (S413D) mutants of NUMB in melanoma cells implicated the involvement of NUMB-S413 phosphorylation in cell migration and invasion, which was independent of NOTCH activation. To determine the clinical relevance of these findings, immunohistochemistry was performed using melanoma tissue microarray, which indicated a strong positive correlation between PLK1 and N-cadherin, a protein required for successful EMT. These findings were supported by TCGA analysis, where expression of high PLK1 with low NUMB or high NOTCH or N-cadherin showed a significant decrease in survival of melanoma patients. Overall, these results suggest a potential role of PLK1 in EMT, migration, and invasion of melanoma cells. Our findings support the therapeutic targeting of PLK1, NUMB, and NOTCH for melanoma management.https://doi.org/10.1038/s41698-023-00493-7 |
| spellingShingle | Gagan Chhabra Chandra K. Singh Mary A. Ndiaye Shengqin Su Carl A. Shirley Nihal Ahmad Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma npj Precision Oncology |
| title | Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma |
| title_full | Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma |
| title_fullStr | Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma |
| title_full_unstemmed | Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma |
| title_short | Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma |
| title_sort | role of plk1 numb notch in epithelial mesenchymal transition in human melanoma |
| url | https://doi.org/10.1038/s41698-023-00493-7 |
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