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Monitoring Epithelial-Mesenchymal Transition of Pancreatic Cancer Cells via Investigation of Mitochondrial Dysfunction.

Methods and protocols (2020-05-01)
Jae Jun Sim, Keun-Yeong Jeong
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Hanks′ Balanced Salt Solution 10x, Without sodium bicarbonate, 10 ×, liquid, sterile-filtered, suitable for cell culture
SAFC
Trypsin-EDTA Solution 10X