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Preferential induction of apoptotic cell death in melanoma cells as compared with normal keratinocytes using a non-thermal plasma torch

机译:使用非热等离子体炬,与正常角质形成细胞相比,可优先诱导黑色素瘤细胞凋亡

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Selective induction of apoptosis in melanoma cells is optimal for therapeutic development. To achieve this goal, a nonthermal helium plasma torch was modified for use on cultured cells in a temperature-controlled environment. Melanoma cells were targeted with this torch (1) in parallel cultures with keratinocytes, (2) in co-culture with keratinocytes and (3) in a soft agar matrix. Melanoma cells displayed high sensitivity to reactive oxygen species generated by the torch and showed a 6-fold increase in cell death compared with keratinocytes. The extent of cell death was compared between melanoma cells and normal human keratinocytes in both short-term (5 min) co-culture experiments and longer assessments of apoptotic cell death (18-24 h). Following a 10 sec plasma exposure there was a 4.9-fold increase in the cell death of melanoma vs. keratinocytes as measured after 24 h at the target site of the plasma beam. When the treatment time was increased to 30 sec, a 98% cell death was reported for melanoma cells, which was 6-fold greater than the extent of cell death in keratinocytes. Our observations further indicate that this preferential cell death is largely due to apoptosis. In addition, we report that this non-thermal plasma torch kills melanoma cells growing in soft agar, suggesting that the plasma torch is capable of inducing melanoma cell death in 3D settings. We demonstrate that the presence of gap junctions may increase the area of cell death, likely due to the "bystander effect" of passing apoptotic signals between cells. Our findings provide a basis for further development of this non-invasive plasma torch as a potential treatment for melanoma.
机译:黑色素瘤细胞中凋亡的选择性诱导是治疗发展的最佳选择。为实现此目标,对非热氦等离子体炬进行了改造,以用于温度受控环境中的培养细胞。用该炬管将黑素瘤细胞作为目标(1)与角质形成细胞平行培养,(2)与角质形成细胞共培养,(3)在软琼脂基质中。黑色素瘤细胞对由火炬产生的活性氧具有很高的敏感性,并且与角质形成细胞相比,细胞死亡增加了6倍。在短期(5分钟)共培养实验和更长的凋亡细胞死亡评估(18-24小时)中,比较了黑素瘤细胞和正常人角质形成细胞之间的细胞死亡程度。在血浆暴露10秒钟后,在等离子束的目标位置24小时后测得,黑色素瘤细胞相对于角质形成细胞的细胞死亡增加了4.9倍。当治疗时间增加到30秒时,黑色素瘤细胞的细胞死亡报告为98%,是角质形成细胞中细胞死亡程度的6倍。我们的观察结果进一步表明,这种优先的细胞死亡很大程度上是由于细胞凋亡。此外,我们报道了这种非热血浆炬可杀死在软琼脂中生长的黑色素瘤细胞,这表明血浆炬能够在3D环境中诱导黑色素瘤细胞死亡。我们证明间隙连接的存在可能会增加细胞死亡的面积,可能是由于细胞之间传递凋亡信号的“旁观者效应”。我们的发现为进一步开发这种无创血浆炬作为黑色素瘤的潜在治疗方法提供了基础。

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