Book of Abstracts - New Frontiers 2022

Abstracts of poster presentations

EFFECT OF SULFORAPHANE ON DOXORUBICIN-INDUCED TOXICITY IN HEK 293 CELLS

B. Boťanská 1 , P. Sovík 2 , M. Barančík 1

1 Centre of Experimental Medicine., Slovak Academy of Sciences, Bratislava, Slovakia; 2 Slovak University of Technology, Faculty of Chemical and Food Technology, Bratislava, Slovakia Introduction: Sulforaphane (SFN) is natural antioxidant found in various cruciferous vegetables, especially in broccoli. Its important functions, including antioxidant, anticancer, and cytoprotective, have been described. SFN is also known as a potential activator of Nrf-2 (Nuclear factor-erythroid factor 2-related factor 2) signaling pathway. Aim: The aim of our study was to study the impact of SFN on Dox-induced effects in human HEK 293 kidney cells. In this, we investigated the influence of both substances on proteins associated with the regulation of redox signaling and autophagy. Methods: Dox and SFN cytotoxicity was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT test), changes in specific proteins were determined by Western blot. Results: We found that SFN administration significantly increased Nrf-2 protein levels, but Dox administration had no significant effect on Nrf-2. Importantly, preincubation of cells with SFN resulted in a significant increase in Nrf-2 levels compared to cells exposed to Dox alone. In case of Keap-1, we found increased protein levels after Dox administration. SFN treatment did not significant influence Dox-induced effects. Next, we found changes in Beclin-1 protein, a key regulator of autophagy. Dox administration, similarly to Keap-1, significantly increased Beclin-1 levels compared to the control group. However, SFN administration had no significant effect on the levels of this protein. Finally, we observed the changes in heat shock proteins (Hsp40 and Hsp60), proteins which are activated in cell responses to stress stimulus. Different changes in levels of these Hsps indicate their specific role in cell responses to SFN and/or Dox administration. While SFN stimulated increase of Hsp40 protein levels, Dox administration had the opposite effect on Hsp40. SFN administration did not significantly influence Dox-induced effects on Hsp40. In contrast to Hsp40, co-administration of SFN with Dox significantly increased Hsp60 protein levels relative to all experimental groups. Conclusions: The obtained results point to a role of Nrf-2 signaling pathway and autophagy in effects of SFN and Dox. Obtained data may help to better understand the interactions between Nrf-2 signaling pathway and autophagy under oxidative stress conditions.

Keywords: sulforaphane; doxorubicin; HEK 293 cells; Nrf2

Funding:This research was funded by VEGA SR grants no. 2/0179/21, 2/0158/20, and grant of Agency for Research and Development APVV-18-0548.

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