TY  - CONF
AU  - Markelić, Milica
AU  - Otašević, Vesna
AU  - Gudelj, Anđelija
AU  - Saksida, Tamara
AU  - Stančić, Ana
AU  - Veličković, Ksenija
AU  - Krstić, Jelena
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6404
AB  - Recently, it has been suggested that nutrient deprivation (ND) may be effective as an adjuvant 
therapy to hepatocellular carcinoma (HCC) cell treatment with sorafenib (Sfb)1. These results 
suggest that ND-mediated priming of HCC cells to Sfb is positively correlated with the p53 status, 
suggesting the essential role of p53 in priming of HCC cells for regulated cell death (RCD). 
Preliminary data indicated morphological signs of ferroptotic RCD, so we aimed to determine whether 
ferroptosis plays a role in the removal of HCC cells in vitro with respect to their p53 status. To 
this end, p53 wild-type (p53WT) and p53 knockout (p53KO) HepG2 cells were grown in growth medium or 
in starvation medium and treated with Sfb or with ferroptosis inducer, Rsl- 3, for 6 h. 
Morphological signs of RCD and nuclear translocation (i.e. activation) of Nrf2, (master regulator 
of ferroptosis-related signalling pathways), as well as protein levels of antioxidative defence 
(AD) enzymes (CAT, CuZnSOD, MnSOD) and ferroptosis-related proteins (GPX4, xCT) were analysed. The 
AD response to Rsl-3 treatment in p53WT cells was similar regardless of nutritional status, as the 
level of all analysed enzymes increased. The response to Sfb was enhanced by ND as CAT and CuZnSOD 
were elevated. p53KO cells responded quite differently, even when treated with Rsl-3, increasing 
only MnSOD. Starved Sfb-treated p53KO cells even decreased expression of AD enzymes. All signs of a 
proferroptotic response examined were present in starved p53WT cells (regardless of treatment): 
decreased nuclear translocation of Nrf2, GPX4, and xCT expression. Nrf2 activation and GPX4 
expression were also decreased in starved p53KO cells (especially upon treatment with Sfb or 
Rsl-3), but accompanied by compensatory overexpressed xCT. These results may be indicative of 
enhanced AD in p53KO cells and may therefore explain, at least in part, their resistance to 
treatment with Sfb+ND which, as presented here, induces ferroptosis in p53WT HepG2 cells.
PB  - Belgrade: Faculty of Chemistry
C3  - Biochemistry in Biotechnology: Serbian Biochemical Society, Twelfth Conference, International scientific meeting; 2023 Sep 21-23; Belgrade, Serbia
T1  - Proferroptotic response to nutrient deprivation in hepatocellular carcinoma cells is related to p53 status
SP  - 86
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6404
ER  - 

@conference{
author = "Markelić, Milica and Otašević, Vesna and Gudelj, Anđelija and Saksida, Tamara and Stančić, Ana and Veličković, Ksenija and Krstić, Jelena",
year = "2023",
abstract = "Recently, it has been suggested that nutrient deprivation (ND) may be effective as an adjuvant 
therapy to hepatocellular carcinoma (HCC) cell treatment with sorafenib (Sfb)1. These results 
suggest that ND-mediated priming of HCC cells to Sfb is positively correlated with the p53 status, 
suggesting the essential role of p53 in priming of HCC cells for regulated cell death (RCD). 
Preliminary data indicated morphological signs of ferroptotic RCD, so we aimed to determine whether 
ferroptosis plays a role in the removal of HCC cells in vitro with respect to their p53 status. To 
this end, p53 wild-type (p53WT) and p53 knockout (p53KO) HepG2 cells were grown in growth medium or 
in starvation medium and treated with Sfb or with ferroptosis inducer, Rsl- 3, for 6 h. 
Morphological signs of RCD and nuclear translocation (i.e. activation) of Nrf2, (master regulator 
of ferroptosis-related signalling pathways), as well as protein levels of antioxidative defence 
(AD) enzymes (CAT, CuZnSOD, MnSOD) and ferroptosis-related proteins (GPX4, xCT) were analysed. The 
AD response to Rsl-3 treatment in p53WT cells was similar regardless of nutritional status, as the 
level of all analysed enzymes increased. The response to Sfb was enhanced by ND as CAT and CuZnSOD 
were elevated. p53KO cells responded quite differently, even when treated with Rsl-3, increasing 
only MnSOD. Starved Sfb-treated p53KO cells even decreased expression of AD enzymes. All signs of a 
proferroptotic response examined were present in starved p53WT cells (regardless of treatment): 
decreased nuclear translocation of Nrf2, GPX4, and xCT expression. Nrf2 activation and GPX4 
expression were also decreased in starved p53KO cells (especially upon treatment with Sfb or 
Rsl-3), but accompanied by compensatory overexpressed xCT. These results may be indicative of 
enhanced AD in p53KO cells and may therefore explain, at least in part, their resistance to 
treatment with Sfb+ND which, as presented here, induces ferroptosis in p53WT HepG2 cells.",
publisher = "Belgrade: Faculty of Chemistry",
journal = "Biochemistry in Biotechnology: Serbian Biochemical Society, Twelfth Conference, International scientific meeting; 2023 Sep 21-23; Belgrade, Serbia",
title = "Proferroptotic response to nutrient deprivation in hepatocellular carcinoma cells is related to p53 status",
pages = "86",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6404"
}

Markelić, M., Otašević, V., Gudelj, A., Saksida, T., Stančić, A., Veličković, K.,& Krstić, J.. (2023). Proferroptotic response to nutrient deprivation in hepatocellular carcinoma cells is related to p53 status. in Biochemistry in Biotechnology: Serbian Biochemical Society, Twelfth Conference, International scientific meeting; 2023 Sep 21-23; Belgrade, Serbia
Belgrade: Faculty of Chemistry., 86.
https://hdl.handle.net/21.15107/rcub_ibiss_6404

Markelić M, Otašević V, Gudelj A, Saksida T, Stančić A, Veličković K, Krstić J. Proferroptotic response to nutrient deprivation in hepatocellular carcinoma cells is related to p53 status. in Biochemistry in Biotechnology: Serbian Biochemical Society, Twelfth Conference, International scientific meeting; 2023 Sep 21-23; Belgrade, Serbia. 2023;:86.
https://hdl.handle.net/21.15107/rcub_ibiss_6404 .

Markelić, Milica, Otašević, Vesna, Gudelj, Anđelija, Saksida, Tamara, Stančić, Ana, Veličković, Ksenija, Krstić, Jelena, "Proferroptotic response to nutrient deprivation in hepatocellular carcinoma cells is related to p53 status" in Biochemistry in Biotechnology: Serbian Biochemical Society, Twelfth Conference, International scientific meeting; 2023 Sep 21-23; Belgrade, Serbia (2023):86,
https://hdl.handle.net/21.15107/rcub_ibiss_6404 .

TY  - JOUR
AU  - Stančić, Ana
AU  - Saksida, Tamara
AU  - Markelić, Milica
AU  - Vučetić, Milica
AU  - Grigorov, Ilijana
AU  - Martinović, Vesna
AU  - Mićanović, Dragica
AU  - Ivanović, Anđelija
AU  - Veličković, Ksenija
AU  - Savić, Nevena
AU  - Otašević, Vesna
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4910
AB  - The main pathological hallmark of diabetes is the loss of functional β-cells. Among several types of β-cell death in diabetes, the
involvement of ferroptosis remains elusive. Therefore, we investigated the potential of diabetes-mimicking factors: high glucose
(HG), proinflammatory cytokines, hydrogen peroxide (H2O2), or diabetogenic agent streptozotocin (STZ) to induce ferroptosis
of β-cells in vitro. Furthermore, we tested the contribution of ferroptosis to injury of pancreatic islets in an STZ-induced
in vivo diabetic model. All in vitro treatments increased loss of Rin-5F cells along with the accumulation of reactive oxygen
species, lipid peroxides and iron, inactivation of NF-E2-related factor 2 (Nrf2), and decrease in glutathione peroxidase 4
expression and mitochondrial membrane potential (MMP). Ferrostatin 1 (Fer-1), ferroptosis inhibitor, diminished the abovestated effects and rescued cells from death in case of HG, STZ, and H2O2 treatments, while failed to increase MMP and to
attenuate cell death after the cytokines’ treatment. Moreover, Fer-1 protected pancreatic islets from STZ-induced injury in
diabetic in vivo model, since it decreased infiltration of macrophages and accumulation of lipid peroxides and increased the
population of insulin-positive cells. Such results revealed differences between diabetogenic stimuli in determining the destiny of
β-cells, emerging HG, H2O2, and STZ, but not cytokines, as contributing factors to ferroptosis and shed new light on an
antidiabetic strategy based on Nrf2 activation. Thus, targeting ferroptosis in diabetes might be a promising new approach for
preservation of the β-cell population. Our results obtained from in vivo study strongly justify this approach.
PB  - London:Hindawi
T2  - Oxidative Medicine and Cellular Longevity
T1  - Ferroptosis as a Novel Determinant of β-Cell Death in Diabetic Conditions
VL  - 2022
DO  - 10.1155/2022/3873420
SP  - 3873420
ER  - 

@article{
author = "Stančić, Ana and Saksida, Tamara and Markelić, Milica and Vučetić, Milica and Grigorov, Ilijana and Martinović, Vesna and Mićanović, Dragica and Ivanović, Anđelija and Veličković, Ksenija and Savić, Nevena and Otašević, Vesna",
year = "2022",
abstract = "The main pathological hallmark of diabetes is the loss of functional β-cells. Among several types of β-cell death in diabetes, the
involvement of ferroptosis remains elusive. Therefore, we investigated the potential of diabetes-mimicking factors: high glucose
(HG), proinflammatory cytokines, hydrogen peroxide (H2O2), or diabetogenic agent streptozotocin (STZ) to induce ferroptosis
of β-cells in vitro. Furthermore, we tested the contribution of ferroptosis to injury of pancreatic islets in an STZ-induced
in vivo diabetic model. All in vitro treatments increased loss of Rin-5F cells along with the accumulation of reactive oxygen
species, lipid peroxides and iron, inactivation of NF-E2-related factor 2 (Nrf2), and decrease in glutathione peroxidase 4
expression and mitochondrial membrane potential (MMP). Ferrostatin 1 (Fer-1), ferroptosis inhibitor, diminished the abovestated effects and rescued cells from death in case of HG, STZ, and H2O2 treatments, while failed to increase MMP and to
attenuate cell death after the cytokines’ treatment. Moreover, Fer-1 protected pancreatic islets from STZ-induced injury in
diabetic in vivo model, since it decreased infiltration of macrophages and accumulation of lipid peroxides and increased the
population of insulin-positive cells. Such results revealed differences between diabetogenic stimuli in determining the destiny of
β-cells, emerging HG, H2O2, and STZ, but not cytokines, as contributing factors to ferroptosis and shed new light on an
antidiabetic strategy based on Nrf2 activation. Thus, targeting ferroptosis in diabetes might be a promising new approach for
preservation of the β-cell population. Our results obtained from in vivo study strongly justify this approach.",
publisher = "London:Hindawi",
journal = "Oxidative Medicine and Cellular Longevity",
title = "Ferroptosis as a Novel Determinant of β-Cell Death in Diabetic Conditions",
volume = "2022",
doi = "10.1155/2022/3873420",
pages = "3873420"
}

Stančić, A., Saksida, T., Markelić, M., Vučetić, M., Grigorov, I., Martinović, V., Mićanović, D., Ivanović, A., Veličković, K., Savić, N.,& Otašević, V.. (2022). Ferroptosis as a Novel Determinant of β-Cell Death in Diabetic Conditions. in Oxidative Medicine and Cellular Longevity
London:Hindawi., 2022, 3873420.
https://doi.org/10.1155/2022/3873420

Stančić A, Saksida T, Markelić M, Vučetić M, Grigorov I, Martinović V, Mićanović D, Ivanović A, Veličković K, Savić N, Otašević V. Ferroptosis as a Novel Determinant of β-Cell Death in Diabetic Conditions. in Oxidative Medicine and Cellular Longevity. 2022;2022:3873420.
doi:10.1155/2022/3873420 .

Stančić, Ana, Saksida, Tamara, Markelić, Milica, Vučetić, Milica, Grigorov, Ilijana, Martinović, Vesna, Mićanović, Dragica, Ivanović, Anđelija, Veličković, Ksenija, Savić, Nevena, Otašević, Vesna, "Ferroptosis as a Novel Determinant of β-Cell Death in Diabetic Conditions" in Oxidative Medicine and Cellular Longevity, 2022 (2022):3873420,
https://doi.org/10.1155/2022/3873420 . .

TY  - CONF
AU  - Otašević, Vesna
AU  - Saksida, Tamara
AU  - Markelić, Milica
AU  - Veličković, Ksenija
AU  - Grigorov, Ilijana
AU  - Stančić, Ana
PY  - 2022
UR  - uri:https://meetings.embo.org/event/22-thiol-oxidation
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5265
AB  - The key pathological event in the development of diabetes is the loss of functional β-cells. We examined here so far unknown the contribution of ferroptosis to β-cell death in diabetic conditions in vivo and in vitro. Thus, male C57BL/6 mice were divided into three groups: diabetic, (strepozotocin-treated 40 mg/kg/5 days), diabetic+ferrostatin-1-treated (Fer-1, 1 mg/kg from day 1-21) and control, while Rin-5F were treated with diabetes-mimicking factors: high glucose (25mM), hydrogen peroxide (75 μM), or streptozotocin (10mM) for 12h. In diabetic mice a significant increase in macrophage infiltration and lipid peroxide accumulation in pancreatic islets, followed by a decrease in an insulin-positive cell population, expression of GPX4, cysteine/glutamate transporter xCT and heme oxygenase 1 were observed. Applied diabetes-mimicking conditions increased death of Rin-5f β-cells, accumulation of reactive oxygen species, lipid peroxides and iron along with a decrease in the activation of transcription factor Nrf2, expression of GPX4 and mitochondrial membrane potential. The use of ferroptosis inhibitor, Fer-1, completely reversed examined ferroptosis-related changes, i.e. had positive effects on both pancreatic islets of diabetic animals and in vitro β-cells. These results show that modulation, i.e. inhibition of ferroptosis in diabetes may be a promising new approach to conserving β-cell populations and treating diabetes.
PB  - Heidelberg: European Molecular Biology Organization (EMBO)
C3  - EMBO workshop: Thiol oxidation in biology: Biochemical mechanisms to physiological outcome; 2022 Oct 8-13
T1  - Inhibition of ferroptosis increases the survival of β-cells
SP  - 52
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5265
ER  - 

@conference{
author = "Otašević, Vesna and Saksida, Tamara and Markelić, Milica and Veličković, Ksenija and Grigorov, Ilijana and Stančić, Ana",
year = "2022",
abstract = "The key pathological event in the development of diabetes is the loss of functional β-cells. We examined here so far unknown the contribution of ferroptosis to β-cell death in diabetic conditions in vivo and in vitro. Thus, male C57BL/6 mice were divided into three groups: diabetic, (strepozotocin-treated 40 mg/kg/5 days), diabetic+ferrostatin-1-treated (Fer-1, 1 mg/kg from day 1-21) and control, while Rin-5F were treated with diabetes-mimicking factors: high glucose (25mM), hydrogen peroxide (75 μM), or streptozotocin (10mM) for 12h. In diabetic mice a significant increase in macrophage infiltration and lipid peroxide accumulation in pancreatic islets, followed by a decrease in an insulin-positive cell population, expression of GPX4, cysteine/glutamate transporter xCT and heme oxygenase 1 were observed. Applied diabetes-mimicking conditions increased death of Rin-5f β-cells, accumulation of reactive oxygen species, lipid peroxides and iron along with a decrease in the activation of transcription factor Nrf2, expression of GPX4 and mitochondrial membrane potential. The use of ferroptosis inhibitor, Fer-1, completely reversed examined ferroptosis-related changes, i.e. had positive effects on both pancreatic islets of diabetic animals and in vitro β-cells. These results show that modulation, i.e. inhibition of ferroptosis in diabetes may be a promising new approach to conserving β-cell populations and treating diabetes.",
publisher = "Heidelberg: European Molecular Biology Organization (EMBO)",
journal = "EMBO workshop: Thiol oxidation in biology: Biochemical mechanisms to physiological outcome; 2022 Oct 8-13",
title = "Inhibition of ferroptosis increases the survival of β-cells",
pages = "52",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5265"
}

Otašević, V., Saksida, T., Markelić, M., Veličković, K., Grigorov, I.,& Stančić, A.. (2022). Inhibition of ferroptosis increases the survival of β-cells. in EMBO workshop: Thiol oxidation in biology: Biochemical mechanisms to physiological outcome; 2022 Oct 8-13
Heidelberg: European Molecular Biology Organization (EMBO)., 52.
https://hdl.handle.net/21.15107/rcub_ibiss_5265

Otašević V, Saksida T, Markelić M, Veličković K, Grigorov I, Stančić A. Inhibition of ferroptosis increases the survival of β-cells. in EMBO workshop: Thiol oxidation in biology: Biochemical mechanisms to physiological outcome; 2022 Oct 8-13. 2022;:52.
https://hdl.handle.net/21.15107/rcub_ibiss_5265 .

Otašević, Vesna, Saksida, Tamara, Markelić, Milica, Veličković, Ksenija, Grigorov, Ilijana, Stančić, Ana, "Inhibition of ferroptosis increases the survival of β-cells" in EMBO workshop: Thiol oxidation in biology: Biochemical mechanisms to physiological outcome; 2022 Oct 8-13 (2022):52,
https://hdl.handle.net/21.15107/rcub_ibiss_5265 .