Nano-pulse stimulation (NPS) is a novel technology to induce cancer apoptosis. In this study, based on the energy-dose effect of NPS, we designed a special NPS sequence (NPSS) with low field intensity. The effectiveness and mechanisms of NPSS on oral cancer therapy were evaluated by cell proliferation assays, microscopic investigation, JC-1 mitochondrial membrane potential assay, tumor inhibition assays, immunohistochemistry (IHC) assay, Ca²⁺, NOS and ROS detection assays, respectively. The results demonstrated that NPSS treatment significantly inhibited oral cancer growth in vitro and in vivo. Furthermore, we found that NPSS treatment induced an obviously apoptosis and mitochondrial membrane potential (ΔΨm) reduction in Cal-27 cells. Notably, further experiments revealed that the mechanisms of crosstalk signaling between NO, ROS and Ca²⁺ involvement in NPSS treatment. In conclusion, this is a proof-of-concept study that provides a potential alternative strategy for developing and applying NPSS in oral cancer therapy.

纳米脉冲刺激（NPS）是一种诱导癌症细胞凋亡的新技术。在这项研究中，基于NPS的能量剂量效应，我们设计了一种低场强的特殊NPS序列（NPSS）。分别通过细胞增殖测定，显微镜研究，JC-1线粒体膜电位测定，肿瘤抑制测定，免疫组织化学（IHC）测定，Ca 2 ⁺，NOS和ROS检测测定来评估NPSS在口腔癌治疗中的有效性和机制。结果表明，治疗NPSS抑制显著口腔癌生长的体外和体内。此外，我们发现NPSS处理在Cal-27细胞中诱导了明显的凋亡和线粒体膜电位（ΔΨm）降低。值得注意的是，进一步的实验表明，NPSS处理中NO，ROS和Ca ²⁺之间的串扰信号传导机制。总之，这是一项概念验证研究，为在口腔癌治疗中开发和应用NPSS提供了潜在的替代策略。