Analytical Determination of Mitochondrial Function of Excised Solid Tumor Homogenates
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Updated: Oct 23, 2025
Analytical Determination of Mitochondrial Function of Excised Solid Tumor Homogenates
Published on: August 6, 2021
Elizabeth R M Zunica1, Christopher L Axelrod2, L Anne Gilmore3
1Integrated Physiology and Molecular Medicine Laboratory, Pennington Biomedical Research Center; Clinical Oncology and Metabolism, Pennington Biomedical Research Center; Department of Nutrition, Case Western Reserve University.
Abstract:
Mitochondria are essential to the onset and progression of cancer through energy production, reactive oxygen species regulation, and macromolecule synthesis. Genetic and functional adaptations of mitochondria to the tumor environment drive proliferative and metastatic potential. The advent of DNA and RNA sequencing removed critical barriers to the evaluation of genetic mediators of tumorigenesis. However, to date, methodological approaches to evaluate tumor mitochondrial function remain elusive and require technical proficiency limiting the feasibility, ultimately diminishing diagnostic and prognostic value in both experimental and clinical settings. Here, we outline a simple and rapid method to quantify rates of oxidative phosphorylation (OXPHOS) and electron transfer (ET) capacity in freshly excised solid tumor homogenates using high-resolution respirometry. The protocol can be reproducibly applied across species and tumor types as well as adapted to evaluate a diversity of mitochondrial ET pathways. Using this protocol, we demonstrate that mice bearing a luminal B mammary cancer exhibit defective nicotinamide adenine dinucleotide-linked respiration and reliance on succinate to generate adenosine triphosphate via OXPHOS.
Insights
Researchers developed a new method to measure mitochondrial function in tumors. This technique revealed defective respiration in mammary cancers, highlighting a reliance on specific pathways for energy production.
Area of Science:
- Biochemistry
- Cancer Biology
- Mitochondrial Biology
Background:
- Mitochondria play a crucial role in cancer development and progression.
- Tumor cells exhibit genetic and functional mitochondrial adaptations.
- Current methods for assessing tumor mitochondrial function are complex and limited.
Purpose of the Study:
- To present a simple, rapid, and reproducible method for quantifying mitochondrial oxidative phosphorylation (OXPHOS) and electron transfer (ET) capacity in solid tumors.
- To establish a protocol applicable across species and tumor types.
- To evaluate mitochondrial function in a specific cancer model.
Main Methods:
- High-resolution respirometry was used to measure OXPHOS and ET capacity in freshly excised solid tumor homogenates.
- The protocol was adapted to assess various mitochondrial ET pathways.
- Mice with luminal B mammary cancer were utilized to test the method.
Main Results:
- The developed protocol is simple, rapid, and reproducible across species and tumor types.
- Mice with luminal B mammary cancer showed impaired nicotinamide adenine dinucleotide-linked respiration.
- These cancer cells demonstrated a reliance on succinate for adenosine triphosphate generation via OXPHOS.
Conclusions:
- A novel, accessible method for assessing tumor mitochondrial function has been established.
- Mitochondrial respiration defects are present in luminal B mammary cancer.
- Understanding these mitochondrial alterations can inform cancer diagnostics and therapeutics.
