摘要
目的:观察纳秒级陡脉冲诱导人卵巢癌细胞SKOV3凋亡的作用及其对细胞内钙离子浓度[Ca2+]i的影响。方法:纳秒级陡脉冲场强10kV/cm,脉宽100ns,频率1Hz,作用5min。SKOV3细胞经纳秒级陡脉冲作用后4h,用透射电子显微镜,流式细胞术检测细胞凋亡。以Fluo-3/AM为细胞内钙离子的荧光指示剂,用激光扫描共聚焦显微镜检测纳秒级陡脉冲对SKOV3细胞[Ca2+]i的影响及[Ca2+]i变化时Ca2+的来源。结果:透射电子显微镜观察到典型的凋亡细胞形态。流式细胞术结果显示,纳秒级陡脉冲主要诱导细胞早期凋亡,早期凋亡率为(22.21±2.71)%,明显高于对照组的(3.04±0.44)%(P<0.01)。纳秒级陡脉冲可明显升高细胞[Ca2+]i(P<0.01),而与细胞外钙离子浓度无关(P>0.05)。结论:纳秒级陡脉冲能够诱导SKOV3细胞凋亡。细胞内钙释放引起钙离子升高,可能是纳秒级陡脉冲诱导肿瘤细胞凋亡的机制之一。
Objective：To investigate the apoptosis and changes of the intracellular calcium concentration [ Ca^2＋ ]i induced by nanosecond steep pulse on human ovarian carcinoma cell line SKOV3 in vitro. Methods： SKOV3 cells were exposed to nanosecond steep pulse （ 10kV/cm, 100ns, 1Hz in 5 minutes）. After 4hours, the apoptosis of SKOV3 cells was detected by transmission electron microscope and flow cytometry. The change of [ Ca^2＋ ] i in SKOV3 cells and the source of Ca^2＋ during [ Ca^2＋ ] i change were observed by laser scanning confocal microscope using Fluo-3/AM as the calcium fluorescent indicator. Results：The typical morphological features of apoptotic cells were observed by transmission electron microscope. Flow cytometry displayed that nanosecond steep pulse mainly induced early apoptosis. Early apoptotic rate of experimental group was （22.21 ±2.71 ）%, signif/cantly higher than that of the control group （ 3.04 ±0.44） % （ P 〈 0.01 ）. The [ Ca^2＋ ] i was markedly increased by treatment with nanosecond steep pulse（ P 〈 0.01 ）, while the increase showed no relationship to the extracellular calcium concentration （ P 〉 0.05 ）. Conclusion： Nanosecond steep pulse can induce apoptosis of SKOV3 cells. The increase of [ Ca^2＋ ]i through release of intracellular calcium may be one of the mechanisms of the apoptosis.
出处
《现代妇产科进展》
CSCD
北大核心
2007年第11期827-831,共5页
Progress in Obstetrics and Gynecology
基金
国家自然科学基金重点项目(No:50637020)
