Hydrogen peroxide in the reactions of cancer cells to cisplatin

• Published in: Biochimica et biophysica acta. General subjects Vol. 1863; no. 4; pp. 692 - 702
• Main Authors: Nerush, A.S., Shсhukina, K.M., Balalaeva, I.V., Orlova, A.G.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2019
• Subjects: Apoptosis; Cancer cell; Cisplatin; drug therapy; dyes; Flow cytometry; fluorescence; Genetically encoded sensor HyPer2; Hydrogen peroxide; membrane potential; mitochondrial membrane; neoplasm cells; phosphatidylserines; Cancer cell; Flow cytometry; Hydrogen peroxide; Genetically encoded sensor HyPer2; Cisplatin; Apoptosis
• ISSN: 0304-4165; 1872-8006; 1872-8006
• DOI: 10.1016/j.bbagen.2019.01.013

Hydrogen peroxide (H2O2) is thought to be one of the key components involved in the responses of tumor cells to chemotherapy. The aim of this study was to reveal the pathways and the phases of cisplatin-induced cell death that are characterized by changes of H2O2 level. The genetically encoded cytosolic fluorescent sensor HyPer2 was used for flow cytometric analysis of the cisplatin-induced changes in H2O2 level in HeLa Kyoto cells. Using a vital dye and the apoptotic markers PE Annexin V or TMRE the pathways and stages of cell death were investigated simultaneously with HyPer2 response. The H2O2 level was studied separately in viable and early apoptotic cells after 12, 18, 24 h's incubation with cisplatin at several concentrations with or without the scavenger of reactive oxygen species NAC. Cisplatin causes dose- and time-dependent increase of H2O2 level in TMRE-positive and PE Annexin V-negative cancer cells. The scavenging of ROS by NAC decreased the H2O2 level and restored cell viability. Н2О2 generation begins in cells that have already lost mitochondrial membrane potential but have not yet externalized phosphatidylserine. Prevention of apoptosis by NAC confirmed the role of H2O2 in apoptosis induction. This is the first time that the sensor HyPer2 has been used in parallel with apoptotic markers and vital dye to demonstrate the role of H2O2 in different stages and types of tumor cell death under chemotherapeutic action. [Display omitted] •Cisplatin causes dose- and time-dependent increase of H2O2 level in early apoptotic and viable cancer cells.•Accumulation of H2O2 in cells begins before the externalization of phosphatidylserine.•Accumulation of H2O2 occurs in cells with lost mitochondrial membrane potential.•Scavenging of H2O2 prevents the cisplatin-induced apoptosis.