@conference{
author = "Despotović, Ana and Harhaji-Trajković, Ljubica and Trajković, Vladimir and Tovilović-Kovačević, Gordana and Zogović, Nevena",
year = "2021",
abstract = "The aim of this study was to investigate the role of necroptosis inhibitor necrostatin-1
(Nec-1) in death of human glioblastoma U251 cells exposed to ascorbic acid (AA), menadione(MD),
and their combination, in vitro. Nec-1 augmented cytotoxicity of AA+MD, and slightly increased
death of MD-treated U251 cells, as assessed by crystal violet (CV) assay. In line with previous, flow
cytometric analysis of annexin/propidium iodide-stained cells showed that Nec-1 triggered cell
death in MD and significantly enhanced ability of AA+MD to increase number of necrotic cells,
substantiating necrosis as the mechanism of U251 cell death induced by combined treatments
– AA+MD, Nec-1+MD, and Nec-1+AA+MD. Further, Nec-1 elevated mitochondrial and cellular
reactive oxygen species (ROS) generated by both MD and AA+MD co-treatment, as assessed
by flow cytometry analysis of MitoSOX- and DHR-stained cells, respectively. N-acetyl cysteine
(NAC), a well-known antioxidant, opposed U251 cell death induced by AA+MD, Nec-1+MD, and
Nec-1+AA+MD, indicating crucial role of oxidative stress in Nec-1-potentiated cytotoxicity of
MD and AA+MD. Also, Nec-1 activated AMP-activated protein kinase (AMPK), and its effector
molecule ULK1 (Ser317) over the level induced by MD and AA+MD, as showed by immunoblot.
AMPK, highly conserved serine/threonine protein kinase, is activated under the conditions of
oxidative stress probably as a consequence of depleted cellular ATP and elevated AMP levels.
This result implies important role of AMPK in necrosis detected in AA+MD-, Nec-1+MD-, and
Nec-1+AA+MD-treated U251 cells. Therefore, it can be concluded that ability of Nec-1 to enhance
cytotoxic potential of AA+MD co-treatment and trigger cytotoxicity of MD is associated with its
capacity to amplify cellular and mitochondrial ROS production, leading to necrosis-like cell
death of U251 cells. Obtained results reveal potential use of Nec-1 as anti-glioblastoma agent,
especially in combination with AA+MD.",
publisher = "Elsevier Inc.",
journal = "Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia",
title = "Necrostatin-1 enhances menadione/ascorbic acid–induced oxidative stress and their cytotoxic potential in human glioblastoma U251 cell line",
doi = "10.1016/j.freeradbiomed.2021.08.081",
pages = "78"
}