Knockdown of Peroxiredoxin V increased the cytotoxicity of non-thermal plasma-treated culture medium to A549 cells

Aging (Albany NY). 2022 May 11;14(9):4000-4013. doi: 10.18632/aging.204063. Epub 2022 May 11.

Abstract

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.

Keywords: Peroxiredoxin V; cytotoxicity; lung cancer; mitogen-activated protein kinase; non-thermal plasma-activated medium.

MeSH terms

  • A549 Cells
  • Apoptosis / genetics
  • Cell Line, Tumor
  • Culture Media
  • Humans
  • Lung Neoplasms* / pathology
  • Peroxiredoxins* / genetics
  • Reactive Oxygen Species / metabolism

Substances

  • Culture Media
  • Reactive Oxygen Species
  • PRDX5 protein, human
  • Peroxiredoxins