Cancer Metabolites Drive Their Survival [Dec. 2, 2025] DOJINDO LABORATORIES

Cancer cell metabolism is shaped not only by genetic alterations but also by microenvironment-derived metabolites that influence cell fate, epigenetic programs, and treatment responses. Tumor cells exploit nutrient-poor and acidic conditions by rerouting metabolic pathways to sustain proliferation and maintain cellular plasticity. Recent studies have revealed that lactate, produced within tumor tissues, stabilizes stem-like states by epigenetically reinforcing proliferative programs, enabling differentiated cancer cells to reacquire self-renewal capacity. Another recent work shows that acetate released from cancer-associated fibroblasts is converted by ACSS2 into acetyl-CoA, activating transcriptional networks that remodel polyamine metabolism and support cancer cell survival in acidic tumor microenvironments. Together, these findings underscore the emerging concept that microenvironmental metabolites act as regulatory signals that shape tumor behavior beyond their traditional role as metabolic substrates.

Lactate controls cancer stemness and plasticity through epigenetic regulation (Cell Metabolism, 2025)
Summary: This study demonstrates that lactate, a metabolic product of cancer cells, controls the balance between cancer stem cells and differentiated cancer cells by suppressing differentiation and promoting dedifferentiation. Through epigenetic regulation of gene activity and metabolic reprogramming, lactate drives tumor growth, therapy resistance, and tumor recurrence, identifying cancer metabolism as a key regulator of tumor cell identity.

Highlighted technique: Human tumor organoids were engineered to express genetically encoded FRET-based metabolic sensors reporting NAD⁺/NADH redox state, intracellular lactate, and glucose dynamics. Time-lapse 3D imaging was analyzed using the authors’ machine-learning-based single-cell tracking software CellPhenTracker, enabling quantitative reconstruction of differentiation and dedifferentiation dynamics at true single-cell resolution.

Related technique Lactate Assay, Glycolysis/OXPHOS Assay

Cancer-associated fibroblast-derived acetate promotes pancreatic cancer development by altering polyamine metabolism via the ACSS2–SP1–SAT1 axis (Nature Cell Biology, 2024)
Summary: In this study, metabolomic analysis revealed that acetate is highly enriched in conditioned media from cancer-associated fibroblasts and in the interstitial fluid of human pancreatic tumors. This CAF-derived acetate is taken up by pancreatic cancer cells and converted into acetyl-CoA by ACSS2, leading to increased histone and SP1 acetylation and enhanced tumor cell survival and growth under acidic tumor microenvironment conditions.

Highlighted technique: To evaluate how pancreatic cancer cells respond to a tumor-mimicking acidic environment with high acetate levels, cell survival and proliferation assays were performed under precisely controlled conditions.Extracellular acetate concentrations were modulated and acetyl-CoA synthesis was inhibited via ACSS2 suppression to functionally assess acetate-dependent stress tolerance in pancreatic cancer cells.

Related technique Cell Proliferation Assay, Cytotoxicity Assay

Metabolic Activity Assays (click to open/close)

Target	Kit & Probes
Glycolysis/Oxidative phosphorylation Assay	Glycolysis/OXPHOS Assay Kit
Lactate mesurement	Lactate Assay Kit-WST
Oxygen consumption rate assay	Extracellular OCR Plate Assay Kit
ATP mesurement	ATP Assay Kit-Luminescence
Mitochondrial membrane potential detection	JC-1 MitoMP Detection Kit, MT-1 MitoMP Detection Kit
High sensitivity glucose uptake assay for plate reader	Glucose Uptake Plate Assay Kit
High sensitivity glucose uptake assay	Glucose Uptake Assay Kit-Blue / Green / Red

Application Note I (click to open/close)
> Inhibition of Mitochondrial Electron Transport Chain

Antimycin stimulation of Jurkat cells was used to evaluate the changes in cellular state upon inhibition of the mitochondrial electron transport chain using a variety of indicators.

The results showed that inhibition of the electron transport chain resulted in (1) a decrease in mitochondrial membrane potential and (2) a decrease in OCR. In addition, (3) the NAD⁺/NADH ratio of the entire glycolytic pathway decreased due to increased metabolism of pyruvate to lactate to maintain the glycolytic pathway, (4) GSH depletion due to increased reactive oxygen species (ROS), and (6) increase in the NADP⁺/NADPH ratio due to decreased NADH required for glutathione biosynthesis were observed.

Application Note II (click to open/close)
> Tracking ROS and Membrane Potential Decline

After HeLa cells were washed with HBSS, co-stained with MitoBright ROS Deep Red and mitochondrial membrane potential staining dye (JC-1: code MT09), and the generated mitochondrial ROS and membrane potential were observed simultaneously. As a result, the decrease in mitochondrial membrane potential and the generation of mitochondrial ROS are simultaneously observed.

＜LEFT: Imaging Conditions＞(Confocal microscopy)
JC-1: Green Ex = 488, Em = 490-520 nm, Red: Ex = 561, Em = 560-600 nm
MitoBright ROS ：Ex = 633 nm, Em = 640-700 nm
Scale bar: 10 μm

＜Right: Examination Conditions＞(Plate Reader)Tecan, Infinite M200 Pro
JC-1: Green Ex=480-490 nm, Em=525-545 nm; Red: Ex= 530-540 nm, Em=585-605 nm
MitoBright ROS: Ex=545-555 nm, Em = 665-685 nm