Research Paper Volume 14, Issue 9 pp 4000—4013
Knockdown of Peroxiredoxin V increased the cytotoxicity of non-thermal plasma-treated culture medium to A549 cells
- 1 Stem Cell and Regenerative Biology Laboratory, College of Life Science and Biotechnology, Heilongjiang Bayi Agricultural University, Daqing 163319, Heilongjiang, China
- 2 Yabian Academy of Agricultural Science, Longjing 1334000, Jilin, China
- 3 Primate Resources Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup-si 56216, Jeonbuk, Republic of Korea
Received: February 6, 2022 Accepted: April 25, 2022 Published: May 11, 2022https://doi.org/10.18632/aging.204063
How to Cite
Copyright: © 2022 Sun et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Administration of non-thermal plasma therapy via the use of plasma-activated medium (PAM) might be a novel strategy for cancer treatment, as it induces apoptosis by increasing reactive oxygen species (ROS) levels. Peroxiredoxin V (PRDX5) scavenges ROS and reactive nitrogen species and is known to regulate several physiological and pathological reactions. However, its role in lung cancer cells exposed to PAM is unknown. Here, we investigated the effect of PRDX5 in PAM-treated A549 lung cancer cells and determined the mechanism underlying its cytotoxicity. Cell culture medium was treated with low temperature plasma at 16.4 kV for 0, 60, 120, or 180 s to develop PAM. PRDX5 was knocked down in A549 cells via transfection with short hairpin RNA targeting PRDX5. Colony formation and wound healing assays, flow cytometry, fluorescence microscopy, and western blotting were performed to detect the effect of PRDX5 knockdown on PAM-treated A549 cells. PAM showed higher cytotoxicity in lung cancer cells than in control cells, downregulated the mitogen-activated protein kinase signaling pathway, and induced apoptosis. PRDX5 knockdown significantly inhibited cell colony formation and migration, increased ROS accumulation, and reduced mitochondrial membrane potential in lung cancer cells. Hence, PRDX5 knockdown combined with PAM treatment represents an effective option for lung cancer treatment.
AKT: Protein kinase B; BAD: BCL2 associated agonist of cell death; BCL2: B cell lymphoma 2; CSCs: cancer stem cells; DHE: dihydroethidium; DMEM: Dulbecco’s modified Eagle’s medium; FBS: Fetal bovine serum; MAPK: mitogen-activated protein kinase; MMP: mitochondrial membrane potential; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide; NAC: N-acetyl cysteine; NSCLC: non-small cell lung cancer; NTP: non-thermal dielectric barrier discharge plasma; p-ERK: phosphorylated extracellular signal-regulated kinase; p-JNK: phosphorylated c-Jun N-terminal kinase; PAM: plasma-activated medium; PARP: poly(ADP-ribose) polymerase; PBS: phosphate-buffered saline; PE: Phycoerythrin and Hoechst; PRDXs: Peroxiredoxins; ROS: reactive oxygen species; SCLC: small cell lung cancer; shRNA: small hairpin RNA; TBS: tris-buffered saline.