METHYLATION OF Odc1 AND Oaz1 GENES IN LEUKEMIA CELLS L1210 AND P388 UNDER THE ACTION OF ORNITHINE DECARBOXYLASE INHIBITORS
DOI:
https://doi.org/10.15407/oncology.2025.01.062Keywords:
ornithine decarboxylase, DNA methylation, odc1 gene, oaz1 gene, DFMO, MGBG, c-MycAbstract
Summary. Elevated ornithine decarboxylase (ODC) activity is known to be associated with the aggressive progression of many tumors. Inhibition of ODC, both at the level of gene expression and enzymatic activity, is considered a promising direction in the development of targeted therapies. Therefore, investigating the methylation status of odc1 and oaz1 gene promoters under the influence of ODC inhibitors may provide insights into the mechanisms of their antitumor action. Objective: to evaluate the methylation levels of odc1 and oaz1 genes, and the protein levels of ODC and c-Myc in L1210 and P388 leukemia cells under the action of synthetic ODC inhibitors. Object and methods: experiments were conducted on mice bearing L1210 and P388 lymphocytic leukemias using synthetic ODC inhibitors. RT²-PCR (EpiTect Methyl II PCR Primer Assay) and Western blotting were applied to assess gene methylation and protein levels. Results: in L1210-bearing mice treated with DFMO+MGBG, the number of tumor cells in ascitic fluid was twofold lower compared to controls (p < 0.0001), with tumor growth inhibition reaching 49.7%. In P388-bearing mice, cell counts were reduced only by 1.3-fold (p < 0.01), with 27.7% tumor growth inhibition. In L1210 cells, DFMO+MGBG increased promoter methylation of odc1 and oaz1 by 1.9- and 2.3-fold, respectively (p < 0.01). In P388 cells, odc1 and oaz1 promoter methylation increased by 2.5- and 2.3-fold, respectively (p < 0.01). The levels of ODC and c-Myc proteins in L1210 cells decreased by 1.8- and 3.1-fold, respectively, and in P388 cells by 1.5- and 1.2-fold compared to controls (p < 0.01). A strong inverse correlation between ODC protein levels and odc1 and oaz1 methylation was observed in L1210 cells (correlation coefficients –0.55 and –0.49, respectively; p < 0.05), while correlations were weaker in P388 cells (–0.41 and –0.37, respectively; p < 0.05). Conclusions: high promoter methylation of odc1 and oaz1 and low levels of c-Myc protein are associated with decreased ODC expression in both leukemia models, which in turn correlates with the enhanced antitumor efficacy of DFMO+MGBG (p < 0.05). These findings suggest that epigenetic regulation of odc1 and oaz1 genes may be a key mechanism underlying the antitumor activity of ODC inhibitors.
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