Decitabine combined with cold atmospheric plasma induces pyroptosis via the ROS/Caspase-3/GSDME signaling pathway in Ovcar5 cells

• Published in: Biochimica et biophysica acta. General subjects Vol. 1868; no. 6; p. 130602
• Main Authors: Du, Liang, Ming, Huiyun, Yan, Zhuna, Chen, Jinwu, Song, Wencheng, Dai, Haiming
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.06.2024
• Subjects: Antimetabolites, Antineoplastic - pharmacology; Caspase 3 - metabolism; caspase-3; caspase-9; cell cycle; Cell Line, Tumor; Cold atmospheric plasma; Decitabine; Decitabine - pharmacology; flow cytometry; Gasdermins; genes; Humans; membrane potential; Membrane Potential, Mitochondrial - drug effects; methylation; mitochondria; mitochondrial membrane; neoplasm cells; nonthermal processing; Ovcar5; oxidoreductases; Plasma Gases - pharmacology; Pyroptosis; Pyroptosis - drug effects; reactive oxygen species; Reactive Oxygen Species - metabolism; ROS; Signal Transduction - drug effects; Western blotting; Decitabine; Pyroptosis; Cold atmospheric plasma; Ovcar5; ROS
• ISSN: 0304-4165; 1872-8006; 1872-8006
• DOI: 10.1016/j.bbagen.2024.130602

High methylation of the DFNA5 gene results in the absence of GSDME, a key protein that mediates pyroptosis, while decitabine demethylates the DFNA5 gene, resulting in high expression of the GSDME protein. Cold atmospheric plasma (CAP) is a novel anti-cancer method that induces tumor cell death. The pyroptosis induced by decitabine in combination with CAP in Ovcar5 cells was evaluated. In particular, mitochondrial membrane potential was estimated by JC-1 staining, dehydrogenase (LDH) release was assessed by ELISA, Annexin V/PI staining was detected by flow cytometry, the cell cycle changes were evaluated using PI staining followed by detection by flow cytometry, and Caspase-9 cleavage, Caspase-3 cleavage and GSDME expression were evaluated by western blot. Decitabine resulted in high expression of the GSDME in Ovcar5 in a concentration-dependent manner and increased tumor cell sensitivity to CAP. CAP induced mitochondrial damage and activated the Caspase-9/Caspase-3 pathway. Therefore, decitabine combined with CAP induced Ovcar5 cell pyroptosis through Caspase-3 mediated GSDME cleavage. Reactive oxygen species (ROS) generated by CAP treatment played an important role in the CAP/decitabine combination-induced production of ROS, activation of Caspase-9/Caspase-3, GSDME cleavage and pyroptosis that ROS scavenger NAC inhibited all these processes. CAP combined with decitabine induced Caspase-3 activation, which cleaved decitabine-upregulated GSDME and ediated pyroptosis. [Display omitted] •Decitabine elevates GSDME protein through demethylation.•Decitabine enhances the sensitivity of ovcar5 cells.•ROS generated by CAP treatment activated the upstream pathway of GSDME-mediated cell death.•Pyroptosis was induced in ovcar5 cells via caspase-9/caspase-3/GSDME signaling pathway.