Effects of folic acid withdrawal on transcriptomic profiles in murine triple-negative breast cancer cell lines

Dieuwertje E Kok; Ciara H O'Flanagan; Michael F Coleman; Zahra Ashkavand; Stephen D Hursting; Sergey A Krupenko

doi:10.1016/j.biochi.2020.04.005

Effects of folic acid withdrawal on transcriptomic profiles in murine triple-negative breast cancer cell lines

Biochimie. 2020 Jun:173:114-122. doi: 10.1016/j.biochi.2020.04.005. Epub 2020 Apr 15.

Authors

Dieuwertje E Kok¹, Ciara H O'Flanagan², Michael F Coleman³, Zahra Ashkavand⁴, Stephen D Hursting⁵, Sergey A Krupenko⁶

Affiliations

¹ Division of Human Nutrition and Health, Wageningen University & Research, the Netherlands. Electronic address: dieuwertje.kok@wur.nl.
² Department of Nutrition, University of North Carolina at Chapel Hill, Chapel Hill, USA. Electronic address: oflanach@gmail.com.
³ Department of Nutrition, University of North Carolina at Chapel Hill, Chapel Hill, USA; Nutrition Research Institute, University of North Carolina at Chapel Hill, Kannapolis, USA. Electronic address: mcoleman@unc.edu.
⁴ Nutrition Research Institute, University of North Carolina at Chapel Hill, Kannapolis, USA. Electronic address: ashkavz@amc.edu.
⁵ Department of Nutrition, University of North Carolina at Chapel Hill, Chapel Hill, USA; Nutrition Research Institute, University of North Carolina at Chapel Hill, Kannapolis, USA; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, USA. Electronic address: hursting@email.unc.edu.
⁶ Department of Nutrition, University of North Carolina at Chapel Hill, Chapel Hill, USA; Nutrition Research Institute, University of North Carolina at Chapel Hill, Kannapolis, USA. Electronic address: sergey_krupenko@unc.edu.

Abstract

We have previously shown that withdrawal of folic acid led to metabolic reprogramming and a less aggressive phenotype in a mouse cell model of triple-negative breast cancer (TNBC). Herein, we evaluate the effects of folic acid withdrawal on transcriptomic profiles in these cells. Murine cell lines were originally derived from a pool of spontaneous mammary tumors grown in MMTV-Wnt1 transgenic mice. Based on their differential molecular characteristics and metastatic potential, these cell lines were previously characterized as non-metastatic epithelial (E-Wnt), non-metastatic mesenchymal (M-Wnt) and metastatic mesenchymal (metM-Wnt^liver) cells. Using custom two-color 180K Agilent microarrays, we have determined gene expression profiles for three biological replicates of each subtype kept on standard medium (2.2 μM folic acid) or folic acid-free medium for 72 h. The analyses revealed that more genes were differentially expressed upon folic acid withdrawal in M-Wnt cells (1884 genes; Benjamini-Hochberg-adjusted P-value <0.05) compared to E-Wnt and metM-Wnt^liver cells (108 and 222 genes, respectively). Pathway analysis has identified that type I interferon signaling was strongly affected by folic acid withdrawal, with interferon-responsive genes consistently being upregulated upon folic acid withdrawal in M-Wnt cells. Of note, repressed interferon signaling has been established as one of the characteristics of aggressive human TNBC, and hence reactivation of this pathway may be a promising therapeutic approach. Overall, while our study indicates that the response to folic acid withdrawal varies by molecular subtype and cellular phenotype, it also underscores the necessity to further investigate one-carbon metabolism as a potential therapeutic means in the treatment of advanced TNBC.

Keywords: Epithelial-to-mesenchymal transition; Folic acid; Interferon signaling; Metastasis; Transcriptomics; Triple-negative breast cancer.

MeSH terms

Animals
Cell Line, Tumor
Culture Media
Epithelial-Mesenchymal Transition
Female
Folic Acid / administration & dosage*
Mice
Mice, Transgenic
Transcriptome*
Triple Negative Breast Neoplasms / metabolism*
Wnt Signaling Pathway

Substances

Culture Media
Folic Acid

Abstract

MeSH terms

Substances

Grants and funding