Late p65 nuclear translocation in glioblastoma cells indicates non-canonical TLR4 signaling and activation of DNA repair genes
Abstract Glioblastoma (GBM) is the most aggressive brain primary malignancy. Toll-like receptor 4 (TLR4) has a dual role in cell fate, promoting cell survival or death depending on the context. Here, we analyzed TLR4 expression in different grades of astrocytoma, and observed increased expression in...
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| Format: | Article |
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Nature Portfolio
2021-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-020-79356-1 |
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| author | Isabele F. Moretti Antonio M. Lerario Marina Trombetta-Lima Paula R. Sola Roseli da Silva Soares Sueli M. Oba-Shinjo Suely K. N. Marie |
| author_facet | Isabele F. Moretti Antonio M. Lerario Marina Trombetta-Lima Paula R. Sola Roseli da Silva Soares Sueli M. Oba-Shinjo Suely K. N. Marie |
| author_sort | Isabele F. Moretti |
| collection | DOAJ |
| description | Abstract Glioblastoma (GBM) is the most aggressive brain primary malignancy. Toll-like receptor 4 (TLR4) has a dual role in cell fate, promoting cell survival or death depending on the context. Here, we analyzed TLR4 expression in different grades of astrocytoma, and observed increased expression in tumors, mainly in GBM, compared to non-neoplastic brain tissue. TLR4 role was investigated in U87MG, a GBM mesenchymal subtype cell line, upon LPS stimulation. p65 nuclear translocation was observed in late phase, suggesting TLR4-non-canonical pathway activation. In fact, components of ripoptosome and inflammasome cascades were upregulated and they were significantly correlated in GBMs of the TCGA-RNASeq dataset. Moreover, an increased apoptotic rate was observed when the GBM-derived U87MG cells were co-treated with LPS and Temozolomide (TMZ) in comparison to TMZ alone. Increased TLR4 immunostaining was detected in nuclei of U87MG cells 12 h after LPS treatment, concomitant to activation of DNA repair genes. Time-dependent increased RAD51, FEN1 and UNG expression levels were confirmed after LPS stimulation, which may contribute to tumor cell fitness. Moreover, the combined treatment with the RAD51 inhibitor, Amuvatinib in combination with, TMZ after LPS stimulation reduced tumor cell viability more than with each treatment alone. In conclusion, our results suggest that stimulation of TLR4 combined with pharmacological inhibition of the DNA repair pathway may be an alternative treatment for GBM patients. |
| first_indexed | 2024-12-18T05:19:37Z |
| format | Article |
| id | doaj.art-dbd539efc27248eda907583955f9ce84 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-18T05:19:37Z |
| publishDate | 2021-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-dbd539efc27248eda907583955f9ce842022-12-21T21:19:42ZengNature PortfolioScientific Reports2045-23222021-01-0111111210.1038/s41598-020-79356-1Late p65 nuclear translocation in glioblastoma cells indicates non-canonical TLR4 signaling and activation of DNA repair genesIsabele F. Moretti0Antonio M. Lerario1Marina Trombetta-Lima2Paula R. Sola3Roseli da Silva Soares4Sueli M. Oba-Shinjo5Suely K. N. Marie6Laboratory of Molecular and Cellular Biology (LIM15), Department of Neurology, Faculdade de Medicina FMUSP, Universidade de Sao PauloDepartment of Internal Medicine, Division of Metabolism, Endocrinology and Diabetes, University of MichiganLaboratory of Molecular and Cellular Biology (LIM15), Department of Neurology, Faculdade de Medicina FMUSP, Universidade de Sao PauloLaboratory of Molecular and Cellular Biology (LIM15), Department of Neurology, Faculdade de Medicina FMUSP, Universidade de Sao PauloLaboratory of Molecular and Cellular Biology (LIM15), Department of Neurology, Faculdade de Medicina FMUSP, Universidade de Sao PauloLaboratory of Molecular and Cellular Biology (LIM15), Department of Neurology, Faculdade de Medicina FMUSP, Universidade de Sao PauloLaboratory of Molecular and Cellular Biology (LIM15), Department of Neurology, Faculdade de Medicina FMUSP, Universidade de Sao PauloAbstract Glioblastoma (GBM) is the most aggressive brain primary malignancy. Toll-like receptor 4 (TLR4) has a dual role in cell fate, promoting cell survival or death depending on the context. Here, we analyzed TLR4 expression in different grades of astrocytoma, and observed increased expression in tumors, mainly in GBM, compared to non-neoplastic brain tissue. TLR4 role was investigated in U87MG, a GBM mesenchymal subtype cell line, upon LPS stimulation. p65 nuclear translocation was observed in late phase, suggesting TLR4-non-canonical pathway activation. In fact, components of ripoptosome and inflammasome cascades were upregulated and they were significantly correlated in GBMs of the TCGA-RNASeq dataset. Moreover, an increased apoptotic rate was observed when the GBM-derived U87MG cells were co-treated with LPS and Temozolomide (TMZ) in comparison to TMZ alone. Increased TLR4 immunostaining was detected in nuclei of U87MG cells 12 h after LPS treatment, concomitant to activation of DNA repair genes. Time-dependent increased RAD51, FEN1 and UNG expression levels were confirmed after LPS stimulation, which may contribute to tumor cell fitness. Moreover, the combined treatment with the RAD51 inhibitor, Amuvatinib in combination with, TMZ after LPS stimulation reduced tumor cell viability more than with each treatment alone. In conclusion, our results suggest that stimulation of TLR4 combined with pharmacological inhibition of the DNA repair pathway may be an alternative treatment for GBM patients.https://doi.org/10.1038/s41598-020-79356-1 |
| spellingShingle | Isabele F. Moretti Antonio M. Lerario Marina Trombetta-Lima Paula R. Sola Roseli da Silva Soares Sueli M. Oba-Shinjo Suely K. N. Marie Late p65 nuclear translocation in glioblastoma cells indicates non-canonical TLR4 signaling and activation of DNA repair genes Scientific Reports |
| title | Late p65 nuclear translocation in glioblastoma cells indicates non-canonical TLR4 signaling and activation of DNA repair genes |
| title_full | Late p65 nuclear translocation in glioblastoma cells indicates non-canonical TLR4 signaling and activation of DNA repair genes |
| title_fullStr | Late p65 nuclear translocation in glioblastoma cells indicates non-canonical TLR4 signaling and activation of DNA repair genes |
| title_full_unstemmed | Late p65 nuclear translocation in glioblastoma cells indicates non-canonical TLR4 signaling and activation of DNA repair genes |
| title_short | Late p65 nuclear translocation in glioblastoma cells indicates non-canonical TLR4 signaling and activation of DNA repair genes |
| title_sort | late p65 nuclear translocation in glioblastoma cells indicates non canonical tlr4 signaling and activation of dna repair genes |
| url | https://doi.org/10.1038/s41598-020-79356-1 |
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