DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines
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 f...
| Main Authors: | , , , , , , , , , , , , , |
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MDPI AG
2022-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/4/2117 |
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| author | Maria Urbanova Verona Buocikova Lenka Trnkova Sabina Strapcova Viera Horvathova Kajabova Emma Barreto Melian Maria Novisedlakova Miroslav Tomas Peter Dubovan Julie Earl Jozef Bizik Eliska Svastova Sona Ciernikova Bozena Smolkova |
| author_facet | Maria Urbanova Verona Buocikova Lenka Trnkova Sabina Strapcova Viera Horvathova Kajabova Emma Barreto Melian Maria Novisedlakova Miroslav Tomas Peter Dubovan Julie Earl Jozef Bizik Eliska Svastova Sona Ciernikova Bozena Smolkova |
| author_sort | Maria Urbanova |
| collection | DOAJ |
| description | 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% O<sub>2</sub>) 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. |
| first_indexed | 2024-03-09T21:45:29Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T21:45:29Z |
| publishDate | 2022-02-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-e3b00744b0f14efcb916cefe3c9bfda62023-11-23T20:20:16ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-02-01234211710.3390/ijms23042117DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell LinesMaria Urbanova0Verona Buocikova1Lenka Trnkova2Sabina Strapcova3Viera Horvathova Kajabova4Emma Barreto Melian5Maria Novisedlakova6Miroslav Tomas7Peter Dubovan8Julie Earl9Jozef Bizik10Eliska Svastova11Sona Ciernikova12Bozena Smolkova13Department of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaDepartment of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaDepartment of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaDepartment of Tumor Biology, Institute of Virology, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaDepartment of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaMolecular 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, SpainOncology Outpatient Clinic, Hospital of the Hospitaller Order of Saint John of God, 814 65 Bratislava, SlovakiaDepartment of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaDepartment of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaMolecular 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, SpainDepartment of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaDepartment of Tumor Biology, Institute of Virology, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaDepartment of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaDepartment of Molecular Oncology, Cancer Research Institute, Biomedical Research Center, Slovak Academy of Sciences, Dubravska Cesta 9, 845 05 Bratislava, SlovakiaDue 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% O<sub>2</sub>) 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.https://www.mdpi.com/1422-0067/23/4/2117PDACinflammationhypoxiaepithelial-to-mesenchymal transitionDNA methylation |
| spellingShingle | Maria Urbanova Verona Buocikova Lenka Trnkova Sabina Strapcova Viera Horvathova Kajabova Emma Barreto Melian Maria Novisedlakova Miroslav Tomas Peter Dubovan Julie Earl Jozef Bizik Eliska Svastova Sona Ciernikova Bozena Smolkova DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines International Journal of Molecular Sciences PDAC inflammation hypoxia epithelial-to-mesenchymal transition DNA methylation |
| title | DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines |
| title_full | DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines |
| title_fullStr | DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines |
| title_full_unstemmed | DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines |
| title_short | DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines |
| title_sort | dna methylation mediates emt gene expression in human pancreatic ductal adenocarcinoma cell lines |
| topic | PDAC inflammation hypoxia epithelial-to-mesenchymal transition DNA methylation |
| url | https://www.mdpi.com/1422-0067/23/4/2117 |
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