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Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo
SIMPLE SUMMARY: Glioblastoma is extremely lethal brain cancer. In our research, we investigated the effect of anti-vimentin nanobody Nb79 on glioblastoma cell invasion. The expression of vimentin in glioma tissues and cells was determined by RT-qPCR. Invasion assay was performed on differentiated gl...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9913279/ https://www.ncbi.nlm.nih.gov/pubmed/36765531 http://dx.doi.org/10.3390/cancers15030573 |
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author | Zottel, Alja Novak, Metka Šamec, Neja Majc, Bernarda Colja, Sara Katrašnik, Mojca Vittori, Miloš Hrastar, Barbara Rotter, Ana Porčnik, Andrej Lah Turnšek, Tamara Komel, Radovan Breznik, Barbara Jovčevska, Ivana |
author_facet | Zottel, Alja Novak, Metka Šamec, Neja Majc, Bernarda Colja, Sara Katrašnik, Mojca Vittori, Miloš Hrastar, Barbara Rotter, Ana Porčnik, Andrej Lah Turnšek, Tamara Komel, Radovan Breznik, Barbara Jovčevska, Ivana |
author_sort | Zottel, Alja |
collection | PubMed |
description | SIMPLE SUMMARY: Glioblastoma is extremely lethal brain cancer. In our research, we investigated the effect of anti-vimentin nanobody Nb79 on glioblastoma cell invasion. The expression of vimentin in glioma tissues and cells was determined by RT-qPCR. Invasion assay was performed on differentiated glioblastoma cell line U-87 MG and stem cell line NCH421k in vitro and in vivo in zebrafish embryos. The effect of Nb79 on expression of EMT biomarkers was determined by Western blot and immunocytochemistry. Our study shows that vimentin is upregulated in glioblastoma tissue compared to lower grade glioma and non-tumour brain tissue. Nb79 reduced glioblastoma cell invasion by up to 64% in vitro and up to 21% in vivo. The tight junction protein ZO-1 had higher expression on the cell membrane, when treated with Nb79 compared to control. In conclusion, our results suggest that anti-vimentin nanobody is a promising tool to target glioblastoma cell invasion. ABSTRACT: Purpose: Glioblastoma (GBM) is the most common primary brain tumour and one of the deadliest cancers. In addition to late diagnosis and inadequate treatment, the extremely low survival rate is also due to the lack of appropriate therapeutic biomarkers and corresponding therapeutic agents. One of the potential therapeutic biomarkers is the intermediate filament vimentin, which is associated with epithelial-mesenchymal transition (EMT). The purpose of this study was to analyse the effect of the anti-vimentin nanobody Nb79 on cell invasion in vitro and in vivo. To further our understanding of the mechanism of action, we investigated the association between Nb79 and EMT in GBM and GBM stem cells by analysing the expression levels of key EMT-related proteins. Methods: The expression of vimentin in glioma tissues and cells was determined by RT-qPCR. An invasion assay was performed on differentiated glioblastoma cell line U-87 MG and stem cell line NCH421k in vitro as well as in vivo in zebrafish embryos. The effect of Nb79 on expression of EMT biomarkers beta-catenin, vimentin, ZEB-1 and ZO1 was determined by Western blot and immunocytochemistry. Results: Our study shows that vimentin is upregulated in glioblastoma tissue compared to lower grade glioma and non-tumour brain tissue. We demonstrated that treatment with Nb79 reduced glioblastoma cell invasion by up to 64% in vitro and up to 21% in vivo. In addition, we found that the tight junction protein ZO-1 had higher expression on the cell membrane, when treated with inhibitory anti-vimentin Nb79 compared to control. Conclusion: In conclusion, our results suggest that anti-vimentin nanobody Nb79 is a promising tool to target glioblastoma cell invasion. |
format | Online Article Text |
id | pubmed-9913279 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-99132792023-02-11 Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo Zottel, Alja Novak, Metka Šamec, Neja Majc, Bernarda Colja, Sara Katrašnik, Mojca Vittori, Miloš Hrastar, Barbara Rotter, Ana Porčnik, Andrej Lah Turnšek, Tamara Komel, Radovan Breznik, Barbara Jovčevska, Ivana Cancers (Basel) Article SIMPLE SUMMARY: Glioblastoma is extremely lethal brain cancer. In our research, we investigated the effect of anti-vimentin nanobody Nb79 on glioblastoma cell invasion. The expression of vimentin in glioma tissues and cells was determined by RT-qPCR. Invasion assay was performed on differentiated glioblastoma cell line U-87 MG and stem cell line NCH421k in vitro and in vivo in zebrafish embryos. The effect of Nb79 on expression of EMT biomarkers was determined by Western blot and immunocytochemistry. Our study shows that vimentin is upregulated in glioblastoma tissue compared to lower grade glioma and non-tumour brain tissue. Nb79 reduced glioblastoma cell invasion by up to 64% in vitro and up to 21% in vivo. The tight junction protein ZO-1 had higher expression on the cell membrane, when treated with Nb79 compared to control. In conclusion, our results suggest that anti-vimentin nanobody is a promising tool to target glioblastoma cell invasion. ABSTRACT: Purpose: Glioblastoma (GBM) is the most common primary brain tumour and one of the deadliest cancers. In addition to late diagnosis and inadequate treatment, the extremely low survival rate is also due to the lack of appropriate therapeutic biomarkers and corresponding therapeutic agents. One of the potential therapeutic biomarkers is the intermediate filament vimentin, which is associated with epithelial-mesenchymal transition (EMT). The purpose of this study was to analyse the effect of the anti-vimentin nanobody Nb79 on cell invasion in vitro and in vivo. To further our understanding of the mechanism of action, we investigated the association between Nb79 and EMT in GBM and GBM stem cells by analysing the expression levels of key EMT-related proteins. Methods: The expression of vimentin in glioma tissues and cells was determined by RT-qPCR. An invasion assay was performed on differentiated glioblastoma cell line U-87 MG and stem cell line NCH421k in vitro as well as in vivo in zebrafish embryos. The effect of Nb79 on expression of EMT biomarkers beta-catenin, vimentin, ZEB-1 and ZO1 was determined by Western blot and immunocytochemistry. Results: Our study shows that vimentin is upregulated in glioblastoma tissue compared to lower grade glioma and non-tumour brain tissue. We demonstrated that treatment with Nb79 reduced glioblastoma cell invasion by up to 64% in vitro and up to 21% in vivo. In addition, we found that the tight junction protein ZO-1 had higher expression on the cell membrane, when treated with inhibitory anti-vimentin Nb79 compared to control. Conclusion: In conclusion, our results suggest that anti-vimentin nanobody Nb79 is a promising tool to target glioblastoma cell invasion. MDPI 2023-01-17 /pmc/articles/PMC9913279/ /pubmed/36765531 http://dx.doi.org/10.3390/cancers15030573 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Zottel, Alja Novak, Metka Šamec, Neja Majc, Bernarda Colja, Sara Katrašnik, Mojca Vittori, Miloš Hrastar, Barbara Rotter, Ana Porčnik, Andrej Lah Turnšek, Tamara Komel, Radovan Breznik, Barbara Jovčevska, Ivana Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo |
title | Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo |
title_full | Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo |
title_fullStr | Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo |
title_full_unstemmed | Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo |
title_short | Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo |
title_sort | anti-vimentin nanobody decreases glioblastoma cell invasion in vitro and in vivo |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9913279/ https://www.ncbi.nlm.nih.gov/pubmed/36765531 http://dx.doi.org/10.3390/cancers15030573 |
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