摘要
为研究陡脉冲电场的凋亡效应,以人肝癌细胞系为实验对象,采用激光扫描共聚焦显微镜观察陡脉冲电场作用下细胞内钙离子浓度的变化,采用流式细胞术Annexin V检测陡脉冲电场诱导细胞凋亡。结果发现,陡脉冲电场能使肝癌细胞内钙离子浓度发生变化,变化的程度与陡脉冲电场的参数和胞外是否有钙有关,使得细胞内钙离子浓度稳态失调或大幅度振荡;流式细胞检测结果证实了陡脉冲电场能有效诱导肝癌细胞凋亡(与对照组比较,P<0.01),并且较低的电压峰值、较宽的脉冲宽度(200V/1.3μs)比较高的电压峰值、较窄的脉冲宽度(600V/100ns)能更有效地(P<0.01)诱导肝癌细胞凋亡。实验结果为陡脉冲杀伤肿瘤细胞的机制和参数选择提供了依据。
Calcium is one of the most important secondly messenger in cells, it plays an important role in apoptosis. Measuring the change of intracellular free calcium concentration （[ Ca2＋ ]i ） is a useful method for apoptosis. Exposing liver cancer cell dyed with Fluo-3/AM to steep pulsed electric field （SPEF）, we observed the change of [ Ca2 ＋ ]i using laser scanning confocal microscope （LSCM） to study the apoptosis effect of SPEF. The experiment results showed that steady state of [ Ca2＋ ]i changed in process of exposing to SPEF, the phenomenon kept up even after canceling SPEF, the changing quantity of [ Ca2＋ ]i had relations with parameter （ peak value, pulse width, et al ） of SPEF and extracellular environment （extracellular free calcium existing or not）. The experiment results of flow cytometry （liver cancer cell dyed with Annexin V-FITC） approved that SPEF could induce apoptosis markedly （ P 〈 0.01）; SPEF with lower voltage （2OOV） and longer width （1.3μs） could induce apoptosis more effectively （P 〈 0.01） than SPEF with higher voltage （600V） and shorter width （100ns）. These experiment results provide possible mechanism and parameter selection basis for tumor treatment using SPEF.
出处
《中国生物医学工程学报》
CAS
CSCD
北大核心
2008年第4期551-556,共6页
Chinese Journal of Biomedical Engineering
基金
国家自然科学基金重点项目(50637020)
国家自然科学基金资助项目(50407007)
高等学校博士学科点专项科研基金项目(20050611020)
重庆市科委重大专项(2005AA5008-3)
关键词
陡脉冲电场
细胞凋亡
细胞内钙离子浓度
流式细胞术
激光扫描共聚焦显微镜
steep pulsed electric field （SPEF）
apoptosis
intracellular free calcium concentration （ [ Ca2＋] i ）
flowcytometry
laser scanning confocal microscope （KSCM）
