钙离子在纳秒级陡脉冲诱导卵巢癌细胞SKOV3凋亡通路中的作用  

Ca^(2+) is an important mediator of nanosecond steep pulse-induced apoptosis in human ovarian cancer SKOV3 cells

作　　者：刘丽娟[1] 赵东阳[2] 王建[2] 姚陈果[2] 孙才新[2] 唐均英[1] 

机构地区：[1]重庆医科大学附属第一医院妇产科,重庆400016 [2]重庆大学电气工程学院高电压与电工新技术教育部重点实验室,重庆400044

出　　处：《南方医科大学学报》2011年第5期772-776,共5页Journal of Southern Medical University

基　　金：国家自然科学基金重点项目(50637020)~~

摘　　要：目的证明纳秒级陡脉冲能够诱导人卵巢癌细胞SKOV3凋亡,且钙离子是其凋亡通路的重要介质。方法用不同参数组的纳秒级陡脉冲处理细胞,Annexin V/PI双染流式细胞仪检测其早期凋亡率及坏死率,用0、25、50、100μmol/L BAP-TA-AM分组螯合细胞内因纳秒级陡脉冲处理而增高的游离Ca2+后,MTT检测细胞存活率,Hoechst33342荧光染色检测细胞凋亡率,Western-blot技术检测caspase12蛋白表达的变化。结果 Annexin V/PI双染流式细胞术结果示:最佳治疗参数为场强90KV/cm;脉宽:100ns;脉冲作用时间:30s;频率:1Hz,其凋亡率最高(60.31±5.67)%,坏死率为(1.35±0.39)%;BAPTA-AM作用过的细胞经纳秒级陡脉冲最佳治疗参数处理后细胞存活率明显高于单用纳秒级陡脉冲处理的细胞(P<0.05),凋亡率也同时降低(P<0.05),caspase-12蛋白表达水平也相应降低(P<0.05)。结论纳秒级陡脉冲能诱导SKOV3细胞凋亡,细胞内游离的钙离子增高是其凋亡通路的重要介质,并有可能介导内质网通路。Objective To explore the role of Ca2＋ in nanosecond steep pulse（NSP）-induced apoptosis of human ovarian carcinoma cell line SKOV3 in vitro.Methods The early apoptotic rate of SKOV3 cells treated with NSP was detected by Annexin V/PI double staining and flow ctytometry.MTT assay was used to detect the viability of the cells pretreated with BAPTA-AM（0,25,50 and 100 μmol/L） chelation for 1 h to increase the intracellular free Ca2＋ prior to NSP exposure,and the cell morphological changes and caspase 12 expression were detected using Hoechst 33342 staining and Western blotting,respectively.Results Flow cytometry showed that NSP induced early apoptosis of SKOV3 cells,and the optimal effect was achieved with the treatment parameter configuration of field strength of 90 kV/cm,pulse width of 100 ns,frequency of 1 Hz,and exposure time of 30 s.The highest early apoptotic rate and necrosis rate was（60.31±5.67） % and（1.35±0.39） %,respectively.Pretreatment with BAPTA-AM chelation prior to NSP exposure significantly increased the cell viability（P0.05） ,and resulted also in lowered apoptosis rate and decreased expression of caspase 12（P0.05） .Conclusion NSP can induce apoptosis in SKOV3 cells.Increased intracellular free Ca2＋ functions as an important mediator in NSP-induced cell apoptosis,which may also involve Ca2＋-mediated endo-plasmic reticulum pathway.

关 键 词：纳秒级陡脉冲 卵巢肿瘤 细胞凋亡 钙离子 

分 类 号：R737.31[医药卫生—肿瘤]