纳秒级陡脉冲电场诱导人黑色素瘤细胞裸鼠皮下移植瘤凋亡作用及机制
Experimental studies on apoptosis of A375 cells of xenografted tumors induced by nanosecond pulsed electric field and its mechanism
杨方黎 1唐均英 1米彦 2姚陈果 2王剑飞2
作者信息
- 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
- 折叠
摘要
目的:观察纳秒级陡脉冲对人黑色素瘤细胞裸鼠皮下移植瘤的诱导凋亡作用和Cagpage-3表达的影响.方法:取雌性4～6 wk龄BALB/c裸鼠20只,建立人黑色素瘤皮下移植瘤模型,设对照组和纳秒级陡脉冲处理组,采用参数为300ns,20 kV/cm,f=1 Hz的脉冲电场处理5 min,每2 d处理1次,共3次,分别观察纳秒级陡脉冲处理组和对照组各时间点瘤体体积大小变化,绘制抑瘤曲线;电镜观察肿瘤超微结构改变;采用免疫荧光检测Caspsse-3蛋白表达;RT-PCR方法检测Caspase-3 mRNA表达;荧光分光光度计测定Ca2+浓度.结果:纳秒级陡脉冲处理组裸鼠瘤体体积明显小于对照组(P<0.01);电镜下可见现典型的细胞凋亡改变;纳秒级陡脉冲处理组Caspase-3蛋白表达和Caspase-3 mRNA表达均较对照组明显增多;Ca2+浓度显著增加.结论:纳秒级陡脉冲电场能够诱导在体人黑色素异体移植瘤细胞产生凋亡.
Abstract
AIM: To investigate the effect of nanosecond steep pulse on the growth and apeptosis in A375 cells of xenografted tumors. METHODS: After establishment of 4 to 6 week-old female nude mouse models carrying human melanoma by subcuta-neous inoculationg, 20 mice were randomly divided into 2 groups: Treatment group and control group. The mice were exposed to nanosecond steep pulse(20 kV/cm, 300 ns, 1 Hz in5 min, three times every 2 d), the changes of the tumor body volume were observed and the inhibition rate was calculated. Caspase-3 was determinated by immunofluorescence and RT-PCR and the apoptesis of A375 cells was detected by transmission electron microscope. The changes of [Ca2+]i in A375 cella were observed by fluorescence spectrophotometer using Fluo-3/AM as the calcium fluorescent indicator. RESULTS: The tumor body volume in treatment group was smaller than that in control group (P<0.01) and typical morphological features of apeptotic cells were observed under transmission electron microscope. Immuno-fluorescence and RT-PCR showed that caspase-3 in treatment group increased compared with that in control group. [Ca2+]i markedly increased by treatment with nanosecond steep pulse (P<0.01). CONCLUSION: Nanosecond steep pulse induces apoptosis of transplanted tumors.
关键词
纳秒级陡脉冲/人黑色素瘤细胞A375/凋亡/Caspase-3表达/Ca2+浓度Key words
nanosecond pulsed electric fild/A375 cells/apop-tosis/caspase-3/concentration of Ca2+引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据