Abstract
Hypoxia is associated with tumor aggressiveness and poor prognosis, including breast cancer. Low oxygen levels induces global genomic hypomethylation and hypermethylation of specific loci in tumor cells. DNA methylation is a reversible epigenetic modification, usually associated with gene silencing, contributing to carcinogenesis and tumor progression. Since the effects of DNA methyltransferase inhibitor are context-dependent and as there is little data comparing their molecular effects in normoxic and hypoxic microenvironments in breast cancer, this study aimed to understand the gene expression profiles and molecular effects in response to treatment with DNA methyltransferase inhibitor in normoxia and hypoxia, using the breast cancer model. For this, a cDNA microarray was used to analyze the changes in the transcriptome upon treatment with DNA methyltransferase inhibitor (5-Aza-2’-deoxycytidine: 5-Aza-2’-dC), in normoxia and hypoxia. Furthermore, immunocytochemistry was performed to investigate the effect of 5-Aza-2’-dC on NF-κB/p65 inflammation regulator subcellular localization and expression, in normoxia and hypoxia conditions. We observed that proinflammatory pathways were upregulated by treatment with 5-Aza-2’-dC, in both conditions. However, treatment with 5-Aza-2’-dC in normoxia showed a greater amount of overexpressed proinflammatory pathways than 5-Aza-2’-dC in hypoxia. In this sense, we observed that the NF-κB expression increased only upon 5-Aza-2’-dC in normoxia. Moreover, nuclear staining for NF-κB and NF-κB target genes upregulation, IL1A and IL1B, were also observed after 5-Aza-2’-dC in normoxia. Our results suggest that 5-Aza-2’-dC induces a greater inflammatory change, at the molecular levels, in normoxic than hypoxic tumor microenvironment. These data may support further studies and expand the understanding of the DNA methyltransferase inhibitor effects in different tumor contexts.
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Acknowledgements
We thank to Brazilian National Cancer Institute Graduate Program in Oncology (PPGO-INCA) for the microarray hardware use, and FAPERJ, CNPq and PPGB/PROEX.
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This work was funded by Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (E-26/010.100944/2018), Conselho Nacional de Desenvolvimento Científico e Tecnológico (302505/2018-0) and Programa de pós-graduação em Biociências/Pró-reitoria de extensão.
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AI performed the experiments and wrote the manuscript draft. AI, RJ, NP, KS and PC performed the experiments and participed in analysis. FA and MA conceived the experiments and revised the manuscript.
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Salviano Soares de Amorim, Í., Rodrigues, J.A., Nicolau, P. et al. 5-Aza-2’-deoxycytidine induces a greater inflammatory change, at the molecular levels, in normoxic than hypoxic tumor microenvironment. Mol Biol Rep 48, 1161–1169 (2021). https://doi.org/10.1007/s11033-020-05931-4
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DOI: https://doi.org/10.1007/s11033-020-05931-4