Science and Technology Production
Medicina - Synergistic apoptotic effect of 2´nitroflavone combined with safingol in murine mammary tumor cells
Congress
Authorship:
Juan Manuel Anselmi Relats ; Leonor P. Roguin ; Mariel Marder ; Julieta Marino ; BLANK, VIVIANA CLAUDIADate:
2021Publishing House and Editing Place:
Ministerio de ciencia y TecnologiaSummary *
Synergistic apoptotic effect of 2´nitroflavone combined with safingol in murine mammary tumor cellsJuan Manuel Anselmi Relats, Leonor Roguin, Mariel Marder, Julieta Marino, Viviana Blank In a previous work, we have demonstrated that the synthetic flavonoid 2´-nitroflavone (2NF) is a potent and selective antitumor agent in vitro and in vivo in a murine LM3 breast cancer model. Sphingosine kinase 1 (SphK1), a lipid kinase overexpressed in some mammary tumor cells, regulates the balance between proapoptotic ceramides and prosurvival sphingosine-1-phosphate (S1P). Furthermore, safingol, a competitive SphK1 inhibitor, prevents catabolism of ceramides, contributing to tumor cell death. It has been reported that certain flavonoids exert antitumor activity through an increment in ceramide levels. Thus, we explored the antiproliferative effect of the simultaneous incubation of 2´NF and safingol in LM3 cells and found that they synergistically inhibited cell proliferation. Based on these results, we studied the apoptotic effect of 2NF in combination with safingol. When we evaluated the expression of bax proapoptotic protein, results obtained by Western blot showed that each compound alone did not modify bax expression, but a fivefold increase was observed after cell incubation with both 2NF and safingol for 24 h (p<0.001). Similarly, results obtained by flow cytometry showed a higher increment in the percentage of hypodiploid cells after simultaneous exposure to both compounds (p<0.001, 24h and p<0.05, 48 h). As recent reports demonstrated that some flavonoids inhibit SphK1 activity interacting directly with the kinase, we performed molecular docking analysis with the docking web server SwissDock. Results obtained showed that safingol and 2NF would bind to different molecular regions of the SphK1.Based on the synergistic antiproliferative and apoptotic effects of 2NF in combination with safingol, we propose that the interaction of both molecules with different sites of SphK1 would favor the generation of apoptotic ceramides by inhibiting the formation of S1P. Further experimental approaches should be performed to confirm a direct interaction of 2NF and SphK1. Information provided by the agent in SIGEVAKey Words
ANTITUMORFLAVONOIDSSYNERGISMSPHINGOSINE KINASE