基础医学与临床 ›› 2012, Vol. 32 ›› Issue (5): 493-499.

• 研究论文 • 上一篇    下一篇

纳秒级陡脉冲可用于治疗裸鼠人恶性黑色素瘤

赵雪,唐均英   

  1. 重庆医科大学一附院妇产科
  • 收稿日期:2011-09-26 修回日期:2012-01-04 出版日期:2012-05-05 发布日期:2012-04-16
  • 通讯作者: 唐均英 E-mail:tjy168168@sina.com
  • 基金资助:
    国家自然科学基金重点项目;教育部博士点基金项目;重庆市科委科技项目

Effect of nanosecond pulsed electric fields on treatment of human melanoma A375 cell xenograft in nude mice

  • Received:2011-09-26 Revised:2012-01-04 Online:2012-05-05 Published:2012-04-16

摘要: 摘要 目的:观察纳秒级陡脉冲(Nanosecond Pulsed Electric Fields, nsPEF)治疗裸鼠皮下人恶性黑色素瘤的效果及其机制。方法:建立裸鼠皮下人黑色素瘤模型,随机分为长期治疗组及对照组各10只,观察肿瘤生长、裸鼠存活率时间,和短期治疗组(=10;分2h、4d组及对照组(分别为4、6和6只),以HE染色检查细胞形态,DNA琼脂糖凝胶电泳、TUNEL法分析细胞凋亡,Western blot法、免疫组织化学方法分析组织肿瘤Bax、Bcl-2的表达。两组均用电场峰值40kV/cm、脉冲宽度200ns、频率1Hz、脉冲次数1000个的nsPEF进行治疗。结果:长期治疗后即刻看到肿瘤区均变为灰白色,表面皮肤均没有出血现象,肿瘤质地变硬;随生存时间延长,肿瘤体积较对照组明显缩小(P<0.01);平均生存时间(天)较对照组(天)明显延长(P<0.01);短期治疗组,4d时坏死区域增多;4d时琼脂糖凝胶电泳显示较对照组有明显“ladder”状分布,凋亡率明显高于对照组(P<0.01),较对照组相比Bax表达明显增高(P<0.01)、而Bcl-2表达明显降低(P<0.01)。结论:nsPEF对裸鼠皮下人黑色素瘤有明显的治疗效果,可能是通过调控Bax、Bcl-2 基因表达诱导细胞凋亡。

关键词: 关键词:纳秒级陡脉冲(nsPEF), 人黑色素瘤, 凋亡, Bax, Bcl-2

Abstract: ABSTRACT Objective:To investigate the effect of nanosecond pulsed electric fields(nsPEF)on human melanoma A375 cell xenograft in nude mice and its possible mechanisms. Methods:Nude mice models were randomly divided into long-term treatment group for observing tumor growth and survival time ( ten mice every group),and short-term treatment group (includes 2h group、4d group and control group,4、6 and 6 mice ,respectively) exposed to the 40kV/cm, 200ns, 1HZ, 1000 pulses .Tumor morphology was observed with HE staining; The cell apoptosis was detected by DNA agarose gel electrophoresis and terminaI deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay; The expression of Bax and Bcl-2 protein was detected by Western blot and SP immunohistochemistry. Results: Tumor turned grey without bleeding spots minutes after long-term treatment,the tumor volume was significantly smaller in treatment group than in control group ( P<0.01),the mean survival time was significantly longer in treatment group()than in control group()(P<0.01).Cell necrosis was found in 4d group;DNA ladder was observed in 4d group;the apoptosis rate of tumor cells was significantly higher in 4d group than control group ( P<0.01);the expression of Bax was significantly higher in 4d group than in control group (P<0.01); the expression of Bcl-2 was significantly lower in 4d group than in control group after short-term treatment( P<0.01). Conclusion: nsPEF may cause melanomas complete remission without recurrence by inducing cell apoptosis under the Bax and Bcl-2 gene regulation mechanism .

Key words: [KEY WORDS] Nanosecond pulsed electric fields(nsPEF), Human melanoma, Apoptosis, Bax, Bcl-2