Nanosecond pulsed electric fields (nsPEFs) applied to cells can induce different biological effects depending on pulse duration and field strength. One known process is the induction of apoptosis whereby nsPEFs are currently investigated as a novel cancer therapy. Another and probably related change is the breakdown of the cytoskeleton. We investigated the elasticity of rat liver epithelial cells WB-F344 in a monolayer using atomic force microscopy (AFM) with respect to the potential of cells to undergo malignant transformation or to develop a potential to metastasize. We found that the elastic modulus of the cells decreased significantly within the first 8 min after treatment with 20 pulses of 100 ns and with a field strength of 20 kV/cm but was still higher than the elasticity of their tumorigenic counterpart WB-ras. AFM measurements and immunofluorescent staining showed that the cellular actin cytoskeleton became reorganized within 5 min. However, both a colony formation assay and a cell migration assay revealed no significant changes after nsPEF treatment, implying that cells seem not to adopt malignant characteristics associated with metastasis formation despite the induced transient changes to elasticity and cytoskeleton that can be observed for up to 1 h.
Non-thermal atmospheric pressure plasma consists of partially ionized gas and contains a range of reactive species including biological active ROS and RNS. There are numerous future applications planned in medicine, e.g. blood coagulation, disinfection and wound care. Recently it has been shown that plasma treatment can have lethal effects on bacteria, whereas eukaryotic cells can be promoted to grow and proliferate. The aim of this study was to investigate the impact of non-thermal plasma on the stimulation of mammalian cells. The cells were treated with the atmospheric pressure plasma jet kINPen09 and it was shown that the effects were dependent on treatment time. While long treatment induced apoptosis, short time plasma treatment increased the proliferation. Therefore the underlying processes needed to be identified in order to modulate the plasmas for future applications. Applying state of the art transcriptomic as well as proteomic approaches (DNA-microarrays and HPLC-MS) we identified several hundred genes and their according proteins which were modulated following non-thermal plasma treatment. Latest software solutions (e.g. IPA, Partek) were applied to disentangle the cellular networks connecting the identified molecules. Qualitative Real-Time Polymerase Chain Reaction was used for verification and revealed up-regulation of signaling molecules (e.g. ERK, MEK and JNK) as well as growth factors and cytokines (e.g. VEGF, FGF and IL-6). Furthermore, applying phosphor-specific western blotting or ELISA techniques we also could proof that plasma treatment activated several signaling cascades and induced the secretion of various cytokines (e.g. IL-6 and IL-8). These results underline the huge potential of plasma for cell modulation with regard to signaling processes involved in cell proliferation and differentiation.
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