Abstract
Cold atmospheric plasma (CAP) is a promising approach in anti-cancer therapy, eliminating cancer cells with high selectivity. However, the molecular mechanisms of CAP action are poorly understood. In this study, we investigated CAP effects on calcium homeostasis in melanoma cells. We observed increased cytoplasmic calcium after CAP treatment, which also occurred in the absence of extracellular calcium, indicating the majority of the calcium increase originates from intracellular stores. Application of previously CAP-exposed extracellular solutions also induced cytoplasmic calcium elevations. A substantial fraction of this effect remained when the application was delayed for one hour, indicating the chemical stability of the activating agent(s). Addition of ryanodine and cyclosporin A indicate the involvement of the endoplasmatic reticulum and the mitochondria. Inhibition of the cytoplasmic calcium elevation by the intracellular chelator BAPTA blocked CAP-induced senescence. This finding helps to understand the molecular influence and the mode of action of CAP on tumor cells.
Highlights
Plasma is ionized gas, which is composed of reactive oxygen (ROS) and nitrogen (RNS) species, charged particles and an optical emission in the UV range
We observed dose-dependent effects on malignant melanoma cells by Cold atmospheric plasma (CAP) generated with the Surface Micro Discharge (SMD) technique[18]
We investigated the effect of CAP on changes in the cytoplasmic Ca2+ concentration in the melanoma cell lines Mel Im and Mel Juso by using the fluorescence dye fura-2 AM
Summary
Plasma is ionized gas, which is composed of reactive oxygen (ROS) and nitrogen (RNS) species, charged particles and an optical emission in the UV range. For the generation of direct plasma, the sample itself serves as an electrode and is directly involved in CAP generation[4,5]. “Hybrid” plasma is produced directly, but in contrast to DBD, the sample does not serve as a counter electrode[6]. CAP treatment for 2 min induced DNA damage and resulted in apoptosis of the melanoma cells. Dose-dependent effects of CAP produced by a SMD device could be observed on squamous head and neck cancer cells, leading to reduction of cell viability and DNA damage[19]. Several studies described CAP-induced anti-cancer effects to be caused by ROS and RNS20,21. Since CAP induces apoptosis and senescence[18] the aim of this study was to investigate the impact of CAP on transient cytoplasmic Ca2+ elevation and downstream events in malignant melanoma cells
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