摘要
为研究陡脉冲电场的凋亡效应,以人肝癌细胞为实验对象,采用激光共聚焦扫描显微镜观察陡脉冲电场作用下细胞内线粒体跨膜电位的实时变化。实验结果表明,陡脉冲电场能使线粒体跨膜电位下降,较高的电压峰值、较窄的脉冲宽度(600 V/100 ns)比较低的电压峰值、较宽的脉冲宽度(200V/1.6μs)能使线粒体跨膜电位更快地下降,并且在停止施加陡脉冲电场后线粒体跨膜电位仍然在不断下降,进而有可能使线粒体跨膜电位崩溃,从而诱导肝癌细胞凋亡,为陡脉冲杀伤肿瘤细胞提供了理论依据。
Mitochondrial is the place of energy storage and supply for cell, on the other hand, it plays an important role in apoptosis, while measuring the change of mitochondrial transmembrane potential （△ψm） is a useful method for apoptosis detection. Exposing liver cancer cell （SMMC-7721） dyed with Rhodamine 123 to steep pulsed electric field （SPEF, pulsed electric field with a very steep rise part and a slow fall part）, this paper observed the change of △ψm using laser confocal scanning microscOpe （LCSM, TCSNT made by LEICA） to study the apoptosis effect of SPEF. Cancer cells were cultured, exposed to SPEF and real-time observed in a homemade rectangle chamber （ 20 mm in length, 10ram in width and 10 mm in height）. Cells were divided into three groups： group A （control group）, group B （test group with SPEF of 200V/1.3μs） and group C （test group with SPEF of 600V/100ns）. Fluorescence pictures were sampled every 1.7s beginning at lmin before SPEF exposing, 10min after SPEF exposing, and fluorescence intensity curves of △ψm were obtained. The experimental results show that △ψm decreases gradually in process of exposing to SPEF, SPEF with higher voltage （600 V） and shorter width （100 ns） can cause a quicker decrease than SPEF with lower voltage （ 200 V） and longer width （ 1.3 μs）. Such phenomenon keeps up even after taking away SPEF, this may lead to cOllapse of △ψm. and induce apoptosis with tremendous possibilities. This experiment supplies possible mechanism for tumor treatment using SPEF.
出处
《高电压技术》
EI
CAS
CSCD
北大核心
2007年第2期79-81,100,共4页
High Voltage Engineering
基金
国家自然科学基金项目(50377046
50407007)
重庆市科委科技项目(2005AA5008-3
2005BB5251)
关键词
陡脉冲电场
凋亡
肝癌细胞
线粒体跨膜电位
激光共聚焦扫描显微镜
steep pulsed electric field （SPEF）
apoptosis
liver cancer cell
mitochondrial transmembrane potential （ △ψm ）
laser confocal scanning microscOpe （LCSM）
