Rethinking Glycolysis/OXPHOS in Cancer [June. 4, 2026]

Cancer cell metabolic states have recently been suggested to require multiparametric assessment, including glycolysis/OXPHOS dependence, mitochondrial abundance and membrane potential, and the utilization of metabolic substrates such as lactate. One study revealed that mitochondrial abundance does not necessarily correlate with OXPHOS dependence in CRC cells. Another showed that lactate is not merely a glycolytic waste product but a reusable mitochondrial fuel that can support OXPHOS and stemness in metastatic breast cancer cells.

Mitochondrial, metabolic and bioenergetic adaptations drive plasticity of colorectal cancer cells and shape their chemosensitivity (Markov, et al., Cell Death & Disease, 2025)
Summary: 
This study demonstrates that cancer cell chemosensitivity cannot be understood solely through glycolytic or OXPHOS dependence. Using eight CRC cell lines, the authors showed that mitochondrial abundance does not necessarily reflect mitochondrial function or metabolic reliance. They further linked mitochondrial quantity, protein density, membrane potential, bioenergetic profiles, and metabolite patterns to drug-specific responses.  

Highlighted technique: 
To functionally classify CRC cell metabolism, the authors quantified mitochondrial membrane potential and mitochondrial ROS by flow cytometry, and measured oxygen consumption and extracellular acidification using Seahorse flux analysis to evaluate respiratory capacity, glycolytic activity, and their balance.

A Glycolysis/OXPHOS assay kit and OCR plate-assay kit provides a low-cost, low-cell-number approach screening for samples before committing to full Seahorse analysis.

Lactate mitochondrial oxidation drives stemness potential in metastatic breast cancer (Zhang, et al., Nature communications, 2025)
Summary: 
In this study, lactate is shown to be not merely a glycolytic byproduct but a mitochondrial fuel source in breast cancer cells undergoing metastasis. In detached cells that survive after loss of adhesion, lactate uptake and oxidation through the CD147/MCT1/LDHB complex support TCA metabolism, OXPHOS, and α-KG-DNMT3B-SOX2-associated stemness potential, linking lactate utilization to metastatic adaptation.

Highlighted technique: 
To examine whether lactate serves as a mitochondrial fuel linked to stemness potential, the authors combined isotope tracing of lactate-derived TCA metabolites with measurements of ATP production, Seahorse-based OCR, and Complex I activity. These metabolic readouts were integrated with tumorsphere formation and SOX2 expression to assess whether lactate oxidation supports OXPHOS and stemness-related sphere-forming capacity.

A mitochondrial fractionation kit enables the collection of intact mitochondria from tissue samples, allowing direct assessment of mitochondrial function, including OCR and Complex I activity. Additionally, intracellular ATP and α-KG can serve as readouts of mitochondrial metabolic changes.