2. Academic Validation
  3. DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines

DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines

  • Int J Mol Sci. 2022 Feb 14;23(4):2117. doi: 10.3390/ijms23042117.
Maria Urbanova 1,  Verona Buocikova 1,  Lenka Trnkova 1,  Sabina Strapcova 2,  Viera Horvathova Kajabova 1,  Emma Barreto Melian 3,  Maria Novisedlakova 4,  Miroslav Tomas 1 5,  Peter Dubovan 1 5,  Julie Earl 3,  Jozef Bizik 1,  Eliska Svastova 2,  Sona Ciernikova 1,  Bozena Smolkova 1
Affiliations

Affiliations

  • 1 Department of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, Slovakia.
  • 2 Department of Tumor Biology, Institute of Virology, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, Slovakia.
  • 3 Molecular Epidemiology and Predictive Tumor Markers Group, Ramón y Cajal Health Research Institute (IRYCIS), Biomedical Research Network in Cancer (CIBERONC), Carretera Colmenar Km 9,100, 28034 Madrid, Spain.
  • 4 Oncology Outpatient Clinic, Hospital of the Hospitaller Order of Saint John of God, 814 65 Bratislava, Slovakia.
  • 5 Department of Surgical Oncology, National Cancer Institute, Slovak Medical University, Klenova 1, 833 10 Bratislava, Slovakia.
PMID: 35216235 DOI: 10.3390/ijms23042117
Abstract

Due to abundant stroma and extracellular matrix, accompanied by lack of vascularization, pancreatic ductal adenocarcinoma (PDAC) is characterized by severe hypoxia. Epigenetic regulation is likely one of the mechanisms driving hypoxia-induced epithelial-to-mesenchymal transition (EMT), responsible for PDAC aggressiveness and dismal prognosis. To verify the role of DNA methylation in this process, we assessed gene expression and DNA methylation changes in four PDAC cell lines. BxPC-3, MIA PaCa-2, PANC-1, and SU.86.86 cells were exposed to conditioned media containing cytokines and inflammatory molecules in normoxic and hypoxic (1% O2) conditions for 2 and 6 days. Cancer Inflammation and Immunity Crosstalk and Human Epithelial to Mesenchymal Transition RT² Profiler PCR Arrays were used to identify top deregulated inflammatory and EMT-related genes. Their mRNA expression and DNA methylation were quantified by qRT-PCR and pyrosequencing. BxPC-3 and SU.86.86 cell lines were the most sensitive to hypoxia and inflammation. Although the methylation of gene promoters correlated with gene expression negatively, it was not significantly influenced by experimental conditions. However, DNA Methyltransferase Inhibitor decitabine efficiently decreased DNA methylation up to 53% and reactivated all silenced genes. These results confirm the role of DNA methylation in EMT-related gene regulation and uncover possible new targets involved in PDAC progression.

Keywords

DNA methylation; PDAC; epithelial-to-mesenchymal transition; hypoxia; inflammation.

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