Hypoxia pp 81-93 | Cite as

Evaluating the Metabolic Impact of Hypoxia on Pancreatic Cancer Cells

  • Divya Murthy
  • Enza Vernucci
  • Gennifer Goode
  • Jaime Abrego
  • Pankaj K. SinghEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1742)


Hypoxia is frequently observed in human cancers and induces global metabolic reprogramming that includes an increase in glucose uptake and glycolysis, alterations in NAD(P)H/NAD(P)+ and intracellular ATP levels, and increased utilization of glutamine as the major precursor for fatty acid synthesis. In this chapter, we describe in detail various physiological assays that have been adopted to study the metabolic shift propagated by exposure to hypoxic conditions in pancreatic cell culture model that includes glucose uptake, glutamine uptake, and lactate release by pancreatic cancer cell lines. We have also elaborated the assays to evaluate the ratio of NAD(P)H/NAD(P)+ and intracellular ATP estimation using the commercially available kit to assess the metabolic state of cancer cells.

Key words

Hypoxia HIF Pancreatic cancer Cancer metabolism Glucose uptake Glutamine uptake Lactate release ATP estimation NADPH/NADP+ ratio 


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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Divya Murthy
    • 1
  • Enza Vernucci
    • 1
  • Gennifer Goode
    • 1
  • Jaime Abrego
    • 1
  • Pankaj K. Singh
    • 1
    • 2
    • 3
    • 4
    Email author
  1. 1.The Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical CenterOmahaUSA
  2. 2.Department of Pathology and MicrobiologyUniversity of Nebraska Medical CenterOmahaUSA
  3. 3.Department of Genetics, Cell Biology and AnatomyUniversity of Nebraska Medical CenterOmahaUSA
  4. 4.Department of Biochemistry and Molecular BiologyUniversity of Nebraska Medical CenterOmahaUSA

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