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Distinct Metabolic Profiling of Invading PANC-1 Pancreatic Cancer Cells

Speaker
  • Senior Investigator, National Institutes for Quantum and Radiological Science and Technology, Japan
    Biography
      Dr. Mayumi Fujita received her PhD from the University of Tokyo, Japan, and has been working at National Institutes for Quantum and Radiological Science and Technology in Japan since 2008. She also joined the lab of Dr. David A. Wink at the National Institute of Health last year and investigated the role of nitric oxide in invading cells. She studies the mechanism of cancer cell invasion and developed a real-time imaging model of invading cells.

    Abstract

    During this webcast Dr. Mayumi Fujita of the National Institutes for Quantum and Radiological Science and Technology in Japan will address the method of real-time imaging of invading cells using the IncuCyte Live-Cell Analysis System, and describe how this model is useful for many studies.

    In previous studies, only 1% of cells in the cultured human pancreatic cancer cell line, PANC-1, were capable of invasion through MatrigelÆ in a transwell invasion assay. This suggested that invasive PANC-1 cells have unique characteristics conferring this phenotype. This was further investigated using a 3D spheroid model of PANC-1, embedded in Matrigel, coupled with IncuCyteÆ Live Cell imaging and analysis to capture the movement of the distinct invading population in real time. The identified, invasive PANC-1 were collected and metabolically characterized by CE-TOFMS, and their metabolic profile compared with whole-culture PANC-1. (Fujita et al. 2017, Cancer Science). The invasive PANC-1 cells were distinct from those of the whole cultured PANC-1, with demonstrated increased consumption of ATP, assumed activation of ATP-generating pathways, and higher arginine utilization by NOS. Although they had higher oxidative stress, the invading cells were also more resistant to it with greater survival upon exposure to H2O2. A reduction of intracellular GSH by BSO inhibited PANC-1 invasiveness. Taken together, these results provide a unique metabolic profile for this invasive PANC-1 cell population, as compared to control cells. Such methodology is readily available for similar assessments of invasive cell phenotypes.

    You will learn:

    • A method will be provided for the real-time imaging of cancer cell invasion from 3D spheroid using the IncuCyteÆ Live-Cell Analysis System.
    • The utility of this model to study cancer cell invasion will be discussed, describing methods of invasion, including that by single cells, collected cells, elongated invasion, or rounded invasion.
    • Important methods information for real-time image capture, identification of invasive phenotypes, and integration with metabolomics analysis.
    • How to easily apply the use of IncuCyte technology and associated applications to their cell line of interest for related studies.

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