Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo
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...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-01-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/15/3/573 |
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| author | Alja Zottel Metka Novak Neja Šamec Bernarda Majc Sara Colja Mojca Katrašnik Miloš Vittori Barbara Hrastar Ana Rotter Andrej Porčnik Tamara Lah Turnšek Radovan Komel Barbara Breznik Ivana Jovčevska |
| author_facet | Alja Zottel Metka Novak Neja Šamec Bernarda Majc Sara Colja Mojca Katrašnik Miloš Vittori Barbara Hrastar Ana Rotter Andrej Porčnik Tamara Lah Turnšek Radovan Komel Barbara Breznik Ivana Jovčevska |
| author_sort | Alja Zottel |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-11T09:50:55Z |
| format | Article |
| id | doaj.art-497c79d38f544060970c1070d82424ff |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-11T09:50:55Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-497c79d38f544060970c1070d82424ff2023-11-16T16:14:09ZengMDPI AGCancers2072-66942023-01-0115357310.3390/cancers15030573Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In VivoAlja Zottel0Metka Novak1Neja Šamec2Bernarda Majc3Sara Colja4Mojca Katrašnik5Miloš Vittori6Barbara Hrastar7Ana Rotter8Andrej Porčnik9Tamara Lah Turnšek10Radovan Komel11Barbara Breznik12Ivana Jovčevska13Medical Centre for Molecular Biology, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, SloveniaMedical Centre for Molecular Biology, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, SloveniaMedical Centre for Molecular Biology, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, SloveniaMedical Centre for Molecular Biology, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, SloveniaDepartment of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, SloveniaDepartment of Neurosurgery, University Medical Centre Ljubljana, 1000 Ljubljana, SloveniaDepartment of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, SloveniaMedical Centre for Molecular Biology, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, SloveniaMedical Centre for Molecular Biology, Institute of Biochemistry and Molecular Genetics, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, SloveniaPurpose: 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.https://www.mdpi.com/2072-6694/15/3/573glioblastomavimentinnanobodyinvasionEMT |
| spellingShingle | Alja Zottel Metka Novak Neja Šamec Bernarda Majc Sara Colja Mojca Katrašnik Miloš Vittori Barbara Hrastar Ana Rotter Andrej Porčnik Tamara Lah Turnšek Radovan Komel Barbara Breznik Ivana Jovčevska Anti-Vimentin Nanobody Decreases Glioblastoma Cell Invasion In Vitro and In Vivo Cancers glioblastoma vimentin nanobody invasion EMT |
| 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 | glioblastoma vimentin nanobody invasion EMT |
| url | https://www.mdpi.com/2072-6694/15/3/573 |
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