纳秒脉冲治疗裸鼠皮下移植瘤抗癌机制研究  

Anti-cancer mechanism of nanosecond Pulsed Electric Fields on xenografts in nude mice

作　　者：彭巧[1] 赵雪[1] 姚陈果[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400030

出　　处：《激光杂志》2013年第2期67-69,共3页Laser Journal

基　　金：国家自然科学基金重点项目(50637020);教育部博士点基金项目(20050611020);重庆市科委科技项目(2005AA5008-3)

摘　　要：本研究探讨了纳秒级陡脉冲(nsPEF)对人恶性黑色素瘤A375细胞裸鼠皮下移植瘤模型可能的抗癌机制。通过用游标卡尺测量裸鼠皮下瘤大小,估算抑瘤率。用琼脂糖凝胶电泳、TUNEL法检测肿瘤细胞在纳秒脉冲电场下的细胞凋亡情况,并用HE染色观察细胞形态变化。通过免疫组化法检测瘤体内微血管密度MVD、微血管形态、血管生长因子VEGF和增殖细胞核抗原PCNA的表达。结果发现nsPEF治疗后4d肿瘤生长抑制率达42.6%,琼脂糖凝胶电泳显示治疗后2h组出现阶梯状分布,治疗后4d组阶梯状分布较之明显,对照组未见阶梯状分布;TUNEL法显示治疗组凋亡率均高于对照组,治疗后4d组凋亡率明显高于2h组(P<0.01)。脉冲处理后见肿瘤区即刻变为灰白色;HE染色发现治疗后2h组肿瘤见少许坏死区域、4d组肿瘤坏死区域增多;免疫组化检测发现治疗后4d组微血管腔闭塞状态、MVD、VEGF及PCNA表达较对照组及治疗后2h组明显增多(P<0.01),而对照组与治疗后2h组比较差异无统计学意义(P>0.05)。结果表明纳秒级陡脉冲可能通过诱导细胞大量凋亡,短时间内降低肿瘤血流量,降低细胞增殖能力并降低肿瘤血管形成和重塑以防止肿瘤再生等方面发挥其抗癌效应。To mveshgate the anticancer eitect oi nanosecond Pulsed Electric Fields InsPEF） on human melanoma A375 cell xenograit in nude mice. Tumor size was detected by vemier caliper. The cell apoptosis was detected by DNA agarose gel electrophoresis and tennina I deoxynucleotidyl transferase - mediated dUTP nick end - labeling （TUNEL） assay;Tumor morphology was observed with HE staining; The expression of microvessel density （1VIVD）, vascular endothelial growth factor （VEGF） and proliferating cell nuclear antigen （PCNA） were detected by SP immunohistochemistry. Results showed that tumor growth inhibition rate was 42.6% at 4d after nsPEF treatment. DNA ladder were observed more typically in 4d group than other two groups; The apoptosis rate of tumor cells in 4dgroup was the most highest than other group detected by TUNEL. Tumor turned grey immediately alter treatment; A tew ot necrosis cells were tound in 2h groupp while numerous necrosis were found in 4d group; The state of blocking of capillary vessel was found significantly in 4d group. The expressions of MVD, VEGF and PCNA were significantly lower in 4d group than the other two groups, while no significant difference were found between 2h group and control group. The result indicated that the inhibitory effects of nsPEF may be related to inducing cell apoptosis, reducing blood flow in a short time, decreasing the angiogenesis and discouraging tumor revascularizatian and tumor cell＇ s proliferative activity.

关 键 词：纳秒级陡脉冲 肿瘤血流量 凋亡 血管形成 增殖 

分 类 号：R454[医药卫生—治疗学]