Volume: 3, 2024
3rd International PhD Student’s Conference at the University of Life Sciences in Lublin, Poland:
ENVIRONMENT – PLANT – ANIMAL – PRODUCT
Abstract number: B001
DOI: https://doi.org/10.24326/ICDSUPL3.B001
Published online: 24 April 2024
ICDSUPL, 3, B001 (2024)
∆M4 anticancer peptide induces drug resistance in A549 cells by Nrf2 and anti-oxidative enzymes induction
Aleksandra Brankiewicz1, 2*, Marcin Zawrotniak3, Ibeth Guevara-Lora2
1 Doctoral School of Exact and Natural Sciences, Jagiellonian University in Cracow, Prof. St. Łojasiewicza 11, 30-348 Cracow, Poland
2 Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Cracow, Gronostajowa 7, 30-387 Cracow, Poland
3 Departament of Comparative Biochemistry and Bioanalytics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Cracow, Gronostajowa 7, 30-387 Cracow, Poland
* Corresponding author: aleksandra.brankiewicz@doctoral.uj.edu.pl
Abstract
The development of chemotherapy resistance in cancer poses a challenge in their treatment. Research on peptide-based drugs (anticancer peptides – ACP) obtained from natural products offers new hope for improving anticancer therapies. The aim of this study is to investigate the response of the A549 non-small lung cancer cell line, frequently used in drug development research, to the cecropin-like peptide ΔM4, a promising anticancer peptide. Peptide-induced pro-oxidative activity in cells was determined by ROS measurement using the NBT reduction assay. Catalase activity was assessed by measuring the decrease in H2O2 absorbance as a time function, while mitochondrial and cytoplasmic superoxide dismutase activities were measured after their separation in native PAGE and subsequent NBT staining. Intracellular concentration of GSSG and GSH was determined using a fluorometric method. Gene expression of selected antioxidative enzymes was examined using real time PCR. The total content of cell Nrf2 was analyzed by Western blot and its translocation to the nucleus was examined by fluorescence microscopy. ∆M4 induced a significant increase in ROS production in A549 cells, which resulted in the activation of antioxidant enzymes such as mitochondrial and cytoplasmic superoxide dismutase and catalase. Gene expression of these enzymes increased after peptide treatment. In parallel, increased levels of GSSG were observed in cells. Gene expression of enzymes involved in the regulation of glutathione oxidation varied depending on the incubation time. These observations confirm the protective response of cells to the pro-oxidant effect of the peptide. The total amount of the Nrf2 transcription factor, the main regulator of cellular antioxidant defense, showed an increasing trend, however, significant translocation of this protein to the nuclei was observed, confirming the defense of A549 cells against the action of the peptide. In conclusion, we propose that the primary defense mechanism of A549 cells against ∆M4 peptide may be the activation of intracellular antioxidant enzymes and the activation of genes under the control of the transcription factor Nrf2, which is largely responsible for resistance in this cell line.
Keywords: ACP, non-small-cell lung cancer, oxidative stress, Nrf2, catalase, superoxide dismutase
How to cite
A. Brankiewicz, M. Zawrotniak, I. Guevara-Lora, 2024. ∆M4 anticancer peptide induces drug resistance in A549 cells by Nrf2 and anti-oxidative enzymes induction. In: 3rd International PhD Student’s Conference at the University of Life Sciences in Lublin, Poland: Environment – Plant – Animal – Product. https://doi.org/10.24326/ICDSUPL3.B001