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Triptolide reverses epithelial-mesenchymal transition in glioma cells via inducing autophagy
BACKGROUND: To observe the effects of triptolide (TP) on the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of glioma cells, and to explore the possible mechanisms of phenotypic changes in EMT. METHODS: The U87 and U251 glioma cell lines were treated TP. The Cell Cou...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
AME Publishing Company
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8422147/ https://www.ncbi.nlm.nih.gov/pubmed/34532441 http://dx.doi.org/10.21037/atm-21-2944 |
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author | Lai, Minfang Liu, Lili Zhu, Long Feng, Wenping Luo, Jilai Liu, Yawei Deng, Shengze |
author_facet | Lai, Minfang Liu, Lili Zhu, Long Feng, Wenping Luo, Jilai Liu, Yawei Deng, Shengze |
author_sort | Lai, Minfang |
collection | PubMed |
description | BACKGROUND: To observe the effects of triptolide (TP) on the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of glioma cells, and to explore the possible mechanisms of phenotypic changes in EMT. METHODS: The U87 and U251 glioma cell lines were treated TP. The Cell Counting Kit-8 (CCK-8) method was used to detect the half-maximal inhibitory concentration (IC(50)) of TP in these two cell lines and the inhibition of cell proliferation at the IC(50) concentration. The wound-healing experiment and Transwell invasion assay were used to detect the cells’ migration and invasion abilities, respectively. Using western blot protocol, the expression levels of the EMT markers were analyzed, and the levels of the autophagy markers were also detected. The pEGFP-C2-LC3B plasmid was transfected into glioma cells, and the effect of TP on autophagy was detected by immunofluorescence. A subcutaneous tumor model in nude mice was established to observe the effect of TP on cell proliferation in vivo, and immunohistochemistry (IHC) was used to detect the expression levels of EMT markers in mouse tumor tissues. RESULTS: TP significantly inhibited the proliferation of U87 and U251 cells in a dose- and time-dependent manner. TP had a significant inhibitory effect on the migration and invasion of U87 and U251 cells. Western blot showed that TP reversed the process of EMT in glioma cells, which was evidenced by the upregulated expression of the epithelial marker E-cadherin, and the downregulated expression of the mesenchymal markers N-cadherin, Vimentin, ZEB1, Snail, and Slug. TP increased autophagy in glioma cells, increased the LC3B II/I ratio, and upregulated Beclin-1 and Atg-7 expression. Immunofluorescence showed that the number of autophagosomes increased significantly after TP was applied to cells. In the nude mouse subcutaneous tumor model, experiments revealed an inhibitory effect of TP on glioma cell proliferation in vivo. IHC confirmed that the expression of E-cadherin was upregulated in mouse tumor tissues, while the expression levels of N-Cadherin and Vimentin were downregulated. CONCLUSIONS: TP can inhibit glioma cell proliferation, migration, and invasion, and reverse EMT progression. The possible mechanism of EMT reversal in glioma cells is that TP induces autophagy. |
format | Online Article Text |
id | pubmed-8422147 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | AME Publishing Company |
record_format | MEDLINE/PubMed |
spelling | pubmed-84221472021-09-15 Triptolide reverses epithelial-mesenchymal transition in glioma cells via inducing autophagy Lai, Minfang Liu, Lili Zhu, Long Feng, Wenping Luo, Jilai Liu, Yawei Deng, Shengze Ann Transl Med Original Article BACKGROUND: To observe the effects of triptolide (TP) on the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of glioma cells, and to explore the possible mechanisms of phenotypic changes in EMT. METHODS: The U87 and U251 glioma cell lines were treated TP. The Cell Counting Kit-8 (CCK-8) method was used to detect the half-maximal inhibitory concentration (IC(50)) of TP in these two cell lines and the inhibition of cell proliferation at the IC(50) concentration. The wound-healing experiment and Transwell invasion assay were used to detect the cells’ migration and invasion abilities, respectively. Using western blot protocol, the expression levels of the EMT markers were analyzed, and the levels of the autophagy markers were also detected. The pEGFP-C2-LC3B plasmid was transfected into glioma cells, and the effect of TP on autophagy was detected by immunofluorescence. A subcutaneous tumor model in nude mice was established to observe the effect of TP on cell proliferation in vivo, and immunohistochemistry (IHC) was used to detect the expression levels of EMT markers in mouse tumor tissues. RESULTS: TP significantly inhibited the proliferation of U87 and U251 cells in a dose- and time-dependent manner. TP had a significant inhibitory effect on the migration and invasion of U87 and U251 cells. Western blot showed that TP reversed the process of EMT in glioma cells, which was evidenced by the upregulated expression of the epithelial marker E-cadherin, and the downregulated expression of the mesenchymal markers N-cadherin, Vimentin, ZEB1, Snail, and Slug. TP increased autophagy in glioma cells, increased the LC3B II/I ratio, and upregulated Beclin-1 and Atg-7 expression. Immunofluorescence showed that the number of autophagosomes increased significantly after TP was applied to cells. In the nude mouse subcutaneous tumor model, experiments revealed an inhibitory effect of TP on glioma cell proliferation in vivo. IHC confirmed that the expression of E-cadherin was upregulated in mouse tumor tissues, while the expression levels of N-Cadherin and Vimentin were downregulated. CONCLUSIONS: TP can inhibit glioma cell proliferation, migration, and invasion, and reverse EMT progression. The possible mechanism of EMT reversal in glioma cells is that TP induces autophagy. AME Publishing Company 2021-08 /pmc/articles/PMC8422147/ /pubmed/34532441 http://dx.doi.org/10.21037/atm-21-2944 Text en 2021 Annals of Translational Medicine. All rights reserved. https://creativecommons.org/licenses/by-nc-nd/4.0/Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0 (https://creativecommons.org/licenses/by-nc-nd/4.0/) . |
spellingShingle | Original Article Lai, Minfang Liu, Lili Zhu, Long Feng, Wenping Luo, Jilai Liu, Yawei Deng, Shengze Triptolide reverses epithelial-mesenchymal transition in glioma cells via inducing autophagy |
title | Triptolide reverses epithelial-mesenchymal transition in glioma cells via inducing autophagy |
title_full | Triptolide reverses epithelial-mesenchymal transition in glioma cells via inducing autophagy |
title_fullStr | Triptolide reverses epithelial-mesenchymal transition in glioma cells via inducing autophagy |
title_full_unstemmed | Triptolide reverses epithelial-mesenchymal transition in glioma cells via inducing autophagy |
title_short | Triptolide reverses epithelial-mesenchymal transition in glioma cells via inducing autophagy |
title_sort | triptolide reverses epithelial-mesenchymal transition in glioma cells via inducing autophagy |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8422147/ https://www.ncbi.nlm.nih.gov/pubmed/34532441 http://dx.doi.org/10.21037/atm-21-2944 |
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