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Monitoring Epithelial–Mesenchymal Transition of Pancreatic Cancer Cells via Investigation of Mitochondrial Dysfunction
In this protocol, we introduced a method of measuring mitochondrial dysfunction to confirm the epithelial–mesenchymal transition (EMT) in pancreatic cancer cells under a hypoxic environment. There are many expertized and complicated methods to verify EMT. However, our methods have indicated that EMT...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7359699/ https://www.ncbi.nlm.nih.gov/pubmed/32349411 http://dx.doi.org/10.3390/mps3020032 |
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author | Sim, Jae Jun Jeong, Keun-Yeong |
author_facet | Sim, Jae Jun Jeong, Keun-Yeong |
author_sort | Sim, Jae Jun |
collection | PubMed |
description | In this protocol, we introduced a method of measuring mitochondrial dysfunction to confirm the epithelial–mesenchymal transition (EMT) in pancreatic cancer cells under a hypoxic environment. There are many expertized and complicated methods to verify EMT. However, our methods have indicated that EMT can be identified by examining changes in reactive oxygen species (ROS) generation and membrane potential in mitochondria. To demonstrate whether the changes in the indicators of mitochondrial dysfunction are correlative to EMT, cell morphology, and expression of E-cadherin and N-cadherin were additionally observed. The results verified that a decrease in membrane potential and an increase in ROS in mitochondria were associated with EMT of pancreatic cancer cells. This protocol would be useful as a basis for providing an additional indicator for changes in the tumor microenvironment of pancreatic cancer cells relating to EMT under a hypoxic environment. |
format | Online Article Text |
id | pubmed-7359699 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-73596992020-08-07 Monitoring Epithelial–Mesenchymal Transition of Pancreatic Cancer Cells via Investigation of Mitochondrial Dysfunction Sim, Jae Jun Jeong, Keun-Yeong Methods Protoc Protocol In this protocol, we introduced a method of measuring mitochondrial dysfunction to confirm the epithelial–mesenchymal transition (EMT) in pancreatic cancer cells under a hypoxic environment. There are many expertized and complicated methods to verify EMT. However, our methods have indicated that EMT can be identified by examining changes in reactive oxygen species (ROS) generation and membrane potential in mitochondria. To demonstrate whether the changes in the indicators of mitochondrial dysfunction are correlative to EMT, cell morphology, and expression of E-cadherin and N-cadherin were additionally observed. The results verified that a decrease in membrane potential and an increase in ROS in mitochondria were associated with EMT of pancreatic cancer cells. This protocol would be useful as a basis for providing an additional indicator for changes in the tumor microenvironment of pancreatic cancer cells relating to EMT under a hypoxic environment. MDPI 2020-04-27 /pmc/articles/PMC7359699/ /pubmed/32349411 http://dx.doi.org/10.3390/mps3020032 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Protocol Sim, Jae Jun Jeong, Keun-Yeong Monitoring Epithelial–Mesenchymal Transition of Pancreatic Cancer Cells via Investigation of Mitochondrial Dysfunction |
title | Monitoring Epithelial–Mesenchymal Transition of Pancreatic Cancer Cells via Investigation of Mitochondrial Dysfunction |
title_full | Monitoring Epithelial–Mesenchymal Transition of Pancreatic Cancer Cells via Investigation of Mitochondrial Dysfunction |
title_fullStr | Monitoring Epithelial–Mesenchymal Transition of Pancreatic Cancer Cells via Investigation of Mitochondrial Dysfunction |
title_full_unstemmed | Monitoring Epithelial–Mesenchymal Transition of Pancreatic Cancer Cells via Investigation of Mitochondrial Dysfunction |
title_short | Monitoring Epithelial–Mesenchymal Transition of Pancreatic Cancer Cells via Investigation of Mitochondrial Dysfunction |
title_sort | monitoring epithelial–mesenchymal transition of pancreatic cancer cells via investigation of mitochondrial dysfunction |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7359699/ https://www.ncbi.nlm.nih.gov/pubmed/32349411 http://dx.doi.org/10.3390/mps3020032 |
work_keys_str_mv | AT simjaejun monitoringepithelialmesenchymaltransitionofpancreaticcancercellsviainvestigationofmitochondrialdysfunction AT jeongkeunyeong monitoringepithelialmesenchymaltransitionofpancreaticcancercellsviainvestigationofmitochondrialdysfunction |