Abstract
Introduction
Pteridines include folate-derived metabolites that have been putatively associated with certain cancers in clinical studies. However, their biological significance in cancer metabolism and role in cancer development and progression remains poorly understood.
Objectives
The purpose of this study was to examine the effects of tumorigenicity on pteridine metabolism by studying a panel of 15 pteridine derivatives using a progressive breast cancer cell line model with and without folic acid dosing.
Methods
The MCF10A progressive breast cancer model, including sequentially derived MCF10A (benign), MCF10AT (premalignant), and MCF10CA1a (malignant) cell lines were dosed with 0, 100, and 250 mg/L folic acid. Pteridines were analyzed in both intracellular and extracellular contexts using an improved high-performance liquid chromatography—tandem mass spectrometry method.
Results
Pteridines were located predominately in the extracellular media. Folic acid dosing increased extracellular levels of pterin, 6-hydroxylumazine, xanthopterin, 6-hydroxymethylpterin, and 6-carboxypterin in a dose-dependent manner. In particular, pterin and 6-hydroxylumazine levels were positively correlated with tumorigenicity upon folate dosing.
Conclusions
Folic acid is a primary driver for pteridine metabolism in human breast cell. Higher folate levels contribute to increased formation and excretion of pteridine derivatives to the extracellular media. In breast cancer, this metabolic pathway becomes dysregulated, resulting in the excretion of certain pteridine derivatives and providing in vitro evidence for the observation of elevated pteridines in the urine of breast cancer patients. Finally, this study reports a novel use of the MCF10A progressive breast cancer model for metabolomics applications that may readily be applied to other metabolites of interest.
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Data availability
The data used in this study are available from the corresponding author upon reasonable request.
Code availability
Not applicable.
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Acknowledgements
The authors would like to thank Barbara Ann Karmanos Cancer Institute for providing the MCA 10 A series of cell lines. We appreciate the assistances of Dr. Paul Nam and Dr. Wenyan Liu on analytical method and instrument troubleshooting. Special thanks are also given to Dr. Yinfa Ma for editing this paper.
Funding
This study was funded by National Institute of Health (NIH), National Cancer Institute, Award No. R03CA219337.
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All authors contributed to the study conception and design. All authors commented on previous versions of the manuscript. LR and ZF conducted experiments and wrote the first draft of the manuscript. CB and HS conceived and designed research, analyzed data, and they also edited this manuscript. All authors read and approved the final manuscript.
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The cell lines, including MCF10A, MCF10AT, and MCF10CA1a.cl1, were provided in October, 2019 by Barbara Ann Karmanos Cancer Instritute (4100 John R, Detroit, Michigan 48201) at Wayne State University with a non-exclusive license agreement.
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Rasmussen, L., Foulks, Z., Burton, C. et al. Establishing pteridine metabolism in a progressive isogenic breast cancer cell model. Metabolomics 18, 2 (2022). https://doi.org/10.1007/s11306-021-01861-9
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DOI: https://doi.org/10.1007/s11306-021-01861-9