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Responses of Solid Tumor Cells in DMEM to Reactive Oxygen Species Generated by Non-Thermal Plasma and Chemically Induced ROS Systems

机译:DMEM中的实体肿瘤细胞对非热等离子体和化学诱导的ROS系统产生的活性氧的响应

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In this study, we assessed the role of different reactive oxygen species (ROS) generated by soft jet plasma and chemical-induced ROS systems with regard to cell death in T98G, A549, HEK293 and MRC5 cell lines. For a comparison with plasma, we generated superoxide anion (O2?), hydroxyl radical (HO·), and hydrogen peroxide (H2O2) with chemicals inside an in vitro cell culture. Our data revealed that plasma decreased the viability and intracellular ATP values of cells and increased the apoptotic population via a caspase activation mechanism. Plasma altered the mitochondrial membrane potential and eventually up-regulated the mRNA expression levels of BAX, BAK1 and H2AX gene but simultaneously down-regulated the levels of Bcl-2 in solid tumor cells. Moreover, a western blot analysis confirmed that plasma also altered phosphorylated ERK1/2/MAPK protein levels. At the same time, using ROS scavengers with plasma, we observed that scavengers of HO· (mannitol) and H2O2 (catalase and sodium pyruvate) attenuated the activity of plasma on cells to a large extent. In contrast, radicals generated by specific chemical systems enhanced cell death drastically in cancer as well as normal cell lines in a dose-dependent fashion but not specific with regard to the cell type as compared to plasma.
机译:在这项研究中,我们评估了T98G,A549,HEK293和MRC5细胞系中由软喷射等离子体和化学诱导的ROS系统产生的不同活性氧(ROS)在细胞死亡方面的作用。为了与血浆进行比较,我们生成了超氧阴离子(O 2 ),羟基自由基(HO·)和过氧化氢(H 2 O 2 )与体外细胞培养物中的化学物质。我们的数据显示血浆通过半胱天冬酶激活机制降低了细胞的活力和细胞内ATP值,并增加了凋亡细胞群。血浆改变了线粒体膜电位,并最终上调了BAX,BAK1和H2AX基因的mRNA表达水平,但同时下调了实体肿瘤细胞中Bcl-2的水平。此外,蛋白质印迹分析证实血浆也改变了磷酸化的ERK1 / 2 / MAPK蛋白水平。同时,使用带有血浆的ROS清除剂,我们观察到HO·(甘露醇)和H 2 O 2 (过氧化氢酶和丙酮酸钠)的清除剂减弱了血浆对细胞的影响很大。相反,在癌症以及正常细胞系中,由特定化学系统产生的自由基以剂量依赖性方式大大提高了癌细胞以及正常细胞系中的细胞死亡,但与血浆相比,其细胞类型没有特异性。

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