纳秒级陡脉冲对人卵巢癌细胞SKOV3凋亡及细胞内Ca~(2+)浓度的影响

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。

The effects of nanosecond steep pulse on apoptosis and the concentration of intracellular calcium of human ovarian carcinoma cell line SKOV3

Objective:To investigate the apoptosis and changes of the intracellular calcium concentrationi 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 5minutes).After 4hours,the apoptosis of SKOV3 cells was detected by transmission electron microscope and flow cytometry.The change of i in SKOV3 cells and the source of Ca2+ during 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)%,significantly higher than that of the control group(3.04±0.44)%(P<0.01).The [Ca2+]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 [Ca2+]i through release of intracellular calcium may be one of the mechanisms of the apoptosis.