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高强度皮秒级脉冲电场通过线粒体介导的途径诱导 HeLa 细胞凋亡。

Intense picosecond pulsed electric fields induce apoptosis through a mitochondrial-mediated pathway in HeLa cells.

机构信息

Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, PR China.

出版信息

Mol Med Rep. 2012 Apr;5(4):981-7. doi: 10.3892/mmr.2012.780. Epub 2012 Feb 3.

DOI:10.3892/mmr.2012.780
PMID:22307872
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3493035/
Abstract

The application of pulsed electric fields (PEF) is emerging as a new technique for tumor therapy. Picosecond pulsed electric fields (psPEF) can be transferred to target deep tissue non-invasively and precisely, but the research of the biological effects of psPEF on cells is limited. Electric theory predicts that intense psPEF will target mitochondria and lead to changes in transmembrane potential, therefore, it is hypothesized that it can induce mitochondrial-mediated apoptosis. HeLa cells were exposed to psPEF in this study to investigate this hypothesis. MTT assay demonstrated that intense psPEF significantly inhibited the proliferation of HeLa cells in a dose-dependent manner. Typical characteristics of apoptosis in HeLa cells were observed, using transmission electron microscopy. Loss of mitochondrial transmembrane potential was explored using laser scanning confocal microscopy with Rhodamine-123 (Rh123) staining. Furthermore, the mitochondrial apoptotic events were also confirmed by western blot analysis for the release of cytochrome C and apoptosis-inducing factor from mitochondria into the cytosol. In addition, activation of caspase-3, caspase-9, upregulation of Bax, p53 and downregulation of Bcl-2 were observed in HeLa cells also indicating apoptosis. Taken together, these results demonstrate that intense psPEF induce cell apoptosis through a mitochondrial-mediated pathway.

摘要

脉冲电场（PEF）的应用正成为肿瘤治疗的一种新技术。皮秒脉冲电场（psPEF）可以非侵入性和精确地传递到目标深层组织，但关于 psPEF 对细胞的生物学效应的研究有限。 电学理论预测，强烈的 psPEF 将靶向线粒体并导致跨膜电位变化，因此，假设它可以诱导线粒体介导的细胞凋亡。本研究中用 psPEF 处理 HeLa 细胞来验证这一假说。MTT 检测表明，强烈的 psPEF 以剂量依赖的方式显著抑制 HeLa 细胞的增殖。透射电子显微镜观察到 HeLa 细胞出现典型的凋亡特征。使用罗丹明 123（Rh123）染色的激光共聚焦扫描显微镜探讨线粒体跨膜电位的丧失。此外，通过线粒体细胞色素 C 和凋亡诱导因子从线粒体释放到细胞质中的 Western blot 分析进一步证实了线粒体凋亡事件。此外，还观察到 caspase-3、caspase-9 的激活，Bax、p53 的上调和 Bcl-2 的下调，这也表明发生了细胞凋亡。总之，这些结果表明强烈的 psPEF 通过线粒体介导的途径诱导细胞凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/8c155a4806f7/MMR-05-04-0981-g12.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/92e03aae37b6/MMR-05-04-0981-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/60c7ec4750f2/MMR-05-04-0981-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/ed1de0e77fdd/MMR-05-04-0981-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/a034220ba7d1/MMR-05-04-0981-g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/1d76b20b8609/MMR-05-04-0981-g07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/8b60926c55c0/MMR-05-04-0981-g09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/8c155a4806f7/MMR-05-04-0981-g12.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/92e03aae37b6/MMR-05-04-0981-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/60c7ec4750f2/MMR-05-04-0981-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/ed1de0e77fdd/MMR-05-04-0981-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/a034220ba7d1/MMR-05-04-0981-g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/1d76b20b8609/MMR-05-04-0981-g07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/8b60926c55c0/MMR-05-04-0981-g09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f86b/3493035/8c155a4806f7/MMR-05-04-0981-g12.jpg

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