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Measurement of Reactive Oxygen Species by Fluorescent Probes in Pancreatic Cancer Cells

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Pancreatic Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1882))

Abstract

Pancreatic cancer is a highly lethal disease and is projected to become the second leading cause of cancer-related death by 2020. Among the different subtypes, pancreatic ductal adenocarcinoma (PDAC) is the most common type of pancreatic cancer. The genetic landscape of PDAC shows nearly ubiquitous mutations of KRAS. However, expression of KRAS somatic mutants alone is insufficient to drive PDAC. Redox deregulation may contribute significantly to KRAS-mediated PDAC. Thus, measurement of cellular reactive oxygen species (ROS) levels is essential to determine how oxidative stress affects mutant KRAS and modulates intracellular signaling pathways leading to the change of cellular functions and the development of PDAC. Here we describe the protocol for comparative measurement of several key forms of ROS, including intracellular and mitochondrial levels of superoxide as well as extracellular H2O2 and general cellular ROS, with oxidation-sensitive fluorescent probes using flow cytometry in pancreatic cancer cells or mutant KRAS transformed cells.

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Acknowledgment

This work is supported by the Start-up Funds from Stony Brook University [W. L], the Pilot Project Grant from the Department of Medicine at Stony Brook University [W. L], a P20 grant from NIH [5P20CA192994-02 to W.L as a coinvestigator], and the National Science Foundation of China [81370957 to Y.L].

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Author notes

  1. Yongde Luo

    Present address: Center BioTherapeutics Ltd. Co., Houston, TX, USA

Authors and Affiliations

  1. Center for Cancer and Metabolism Research, Institute for Life Science, Wenzhou University, Zhejiang, China

    Yongde Luo

  2. School of Pharmaceutical Science, Wenzhou Medical University, Zhejiang, China

    Yongde Luo

  3. Proteomics and Nanotechnology Laboratory, Center for Cancer and Stem Cell Biology, Institute of Biosciences and Technology, Texas A&M University Health Science Center, Houston, TX, USA

    Yongde Luo

  4. Division of Gastroenterology and Hepatology, Department of Medicine, Stony Brook University School of Medicine, Stony Brook, NY, USA

    Dan Wang & Weiqin Lu

  5. Division of Medical Oncology, Department of Medicine, Duke Cancer Institute, Duke University, Durham, NC, USA

    James L. Abbruzzese

Authors
  1. Yongde Luo
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  2. Dan Wang
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  3. James L. Abbruzzese
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  4. Weiqin Lu
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Corresponding authors

Correspondence to Yongde Luo or Weiqin Lu .

Editor information

Editors and Affiliations

  1. Department of Pathology and Cell Biology, Columbia University Medical Center, New York, NY, USA

    Gloria H. Su

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Luo, Y., Wang, D., Abbruzzese, J.L., Lu, W. (2019). Measurement of Reactive Oxygen Species by Fluorescent Probes in Pancreatic Cancer Cells. In: Su, G. (eds) Pancreatic Cancer. Methods in Molecular Biology, vol 1882. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8879-2_19

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  • DOI: https://doi.org/10.1007/978-1-4939-8879-2_19

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8878-5

  • Online ISBN: 978-1-4939-8879-2

  • eBook Packages: Springer Protocols

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