Department of Biological Sciences, University at Buffalo, Buffalo, NY, USA.
Cancer Biol Ther. 2012 Nov;13(13):1299-306. doi: 10.4161/cbt.21787. Epub 2012 Aug 16.
Selective induction of apoptosis in melanoma cells is optimal for therapeutic development. To achieve this goal, a non-thermal 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.
选择性诱导黑色素瘤细胞凋亡对于治疗开发是最佳的。为了实现这一目标,我们对非热氦气等离子体火炬进行了改造,使其可在温度可控的环境中用于培养细胞。我们使用该火炬(1)在与角质形成细胞的平行培养物中,(2)在与角质形成细胞的共培养物中,以及(3)在软琼脂基质中靶向黑色素瘤细胞。与角质形成细胞相比,黑素瘤细胞对火炬产生的活性氧具有高敏感性,并显示细胞死亡增加了 6 倍。在短期(5 分钟)共培养实验和对凋亡性细胞死亡(18-24 小时)的更长评估中,比较了黑色素瘤细胞与正常人类角质形成细胞之间的细胞死亡程度。在等离子体束的靶位处,与角质形成细胞相比,暴露于等离子体 10 秒后,黑色素瘤细胞的细胞死亡增加了 4.9 倍。当处理时间延长至 30 秒时,报道了黑色素瘤细胞的 98%细胞死亡,这比角质形成细胞的细胞死亡程度高 6 倍。我们的观察结果进一步表明,这种优先的细胞死亡主要归因于细胞凋亡。此外,我们报告说,这种非热等离子体火炬可杀死在软琼脂中生长的黑色素瘤细胞,这表明等离子体火炬能够在 3D 环境中诱导黑色素瘤细胞死亡。我们证明了缝隙连接的存在可能会增加细胞死亡区域,这可能是由于凋亡信号在细胞之间传递的“旁观者效应”所致。我们的发现为进一步开发这种非侵入性等离子体火炬作为黑色素瘤的潜在治疗方法提供了依据。