Abstract
Cold atmospheric plasma (CAP) has been reported to have strong anticancer effects in vitro and in vivo. CAP has been known to induce apoptosis in most cancer cells by treatment to cells using direct and indirect treatment methods. There are many reports of apoptosis pathways induced by CAP, but for indirect treatment, there is still a lack of fundamental research on how CAP can cause apoptosis in cancer cells. In this study, we applied an indirect treatment method to determine how CAP can induce cancer cell death. First, plasma-activated medium (PAM) was produced by a 2.45 GHz microwave-excited atmospheric pressure plasma jet (ME-APPJ). Next, the amounts of various reactive species in the PAM were estimated using colorimetric methods. The concentration of NO2– and H2O2 in PAM cultured with cancer cells was measured, and intracellular reactive oxidative stress (ROS) changes were observed using flow cytometry. When PAM was incubated with A549 lung cancer cells, there was little change in NO2– concentration, but the concentration of H2O2 gradually decreased after 30 min. While the intracellular ROS of A549 cells was rapidly increased at 2 hours, there was no significant change in that of PAM-treated normal cells. Furthermore, PAM had a significant cytotoxic effect on A549 cells but had little effect on normal cell viability. In addition, using flow cytometry, we confirmed that apoptosis of A549 cells occurred following flow cytometry and western blot analysis. These results suggest that among various reactive species produced by PAM, hydrogen peroxide plays a key role in inducing cancer cell apoptosis.
Highlights
Anticancer therapy using cold atmospheric plasma (CAP) is a rapidly developing field through collaboration with physics, biology, chemistry, and medicine [1,2,3,4,5,6]
The present study investigated the effects of NO2– and H2O2 in the plasma-activated medium (PAM) with A549 human lung cancer cells
These results suggest that hydrogen peroxide in the PAM influenced the increase of the intracellular reactive oxidative stress (ROS) level in A549 cells
Summary
Anticancer therapy using cold atmospheric plasma (CAP) is a rapidly developing field through collaboration with physics, biology, chemistry, and medicine [1,2,3,4,5,6]. CAP contains highly reactive oxygen and nitrogen species (RONS), and it can induce cell death by causing oxidative damage to cancer cells. Direct treatment can induce strong cell death by reaching the plasma species to the cells. Indirect treatment is a method of inducing RONS in a solution by treating plasma to a medium or buffer. It causes less cell death than direct treatment, it has minimal effects on normal cells, is easy to process and store plasmagenerated RONS, and can be treated on large areas. Plasma-activated medium (PAM) generated by indirect treatment can be a good candidate for biomedical applications such as cancer therapy [23]
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