不可逆电穿孔消融技术治疗兔肌肉VX_2移植瘤的研究  

Irreversible electroporation ablation technique in treatment of muscular VX_2 xenografts in rabbits

作　　者：宁周雨[1] 王鹏[1] 陈颢[1] 陈其文[1] 徐立涛[1] 庄丽萍[1] 宋利斌[1] 高嵩[1] 姜峰 孟志强[1] 

机构地区：[1]复旦大学附属肿瘤医院中西医结合科,复旦大学上海医学院肿瘤学系,上海200032 [2]潍坊诸城中医医院肿瘤科,山东潍坊262200

出　　处：《肿瘤》2016年第4期396-405,共10页Tumor

摘　　要：目的:通过观察不可逆电穿孔(irreversible electroporation,IRE)消融技术应用于治疗兔肌肉VX_2移植瘤的疗效及安全性,以期为该技术应用于临床提供可靠的实验依据。方法:建立兔肌肉VX_2移植瘤模型(10只),随机分为IRE治疗组(8只)和对照组(2只,不予以IRE治疗)。分别于IRE治疗的当日处死对照组的兔子、治疗后第1、3、5和14天时分批处死IRE治疗组的兔子(每组2只),同时收集血浆和血清;肿瘤组织行HE染色,并采用免疫组织化学法检测Bcl-2、热休克蛋白70(heat shock protein 70,HSP70)和血管内皮生长因子(vascular endothelial growth factor,VEGF)在肌肉VX_2移植瘤中的表达情况;应用TUNEL法检测移植瘤组织中细胞凋亡的情况;ELISA法检测血浆中caspase 3、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和VEGF的分泌情况,以及血清中实验兔肝肾功能指标、肌酸激酶-MB同功酶(creatine kinase-MB isoenzyme,CK-MB)及心肌肌钙蛋白Ⅰ(cardiac troponinⅠ,CTnⅠ)的表达水平。结果:HE染色结果显示,IRE治疗组中治疗区域和正常组织区域边界清晰,IRE治疗后第3天时,移植瘤组织的边缘区可见大量红细胞以及炎性细胞的浸润,第14天时术区肌肉组织出现纤维化。ELISA法检测结果显示,IRE能有效地增加兔子机体中caspase 3和TNF-α的分泌水平(P值均<0.01),而降低VEGF的分泌水平。同时IRE治疗后,能够降低术区肿瘤组织中Bcl-2和VEGF的阳性表达率(P值均<0.01),但HSP70的表达水平不一。TUNEL检测结果显示,IRE治疗组移植瘤组织中细胞的凋亡率明显增加。术后实验兔的肾功能未见明显异常,谷草转氨酶(aspartate aminotransferase,AST)、谷丙转氨酶(alanine transaminase,ALT)以及CK-MB的表达水平有不同程度的升高,但CTnⅠ的水平未见升高。结论:IRE能够通过诱导细胞凋亡、产生特异的抗肿瘤免疫效应及抑制血管生成来抑制肌肉移植瘤的生长,是一种安全且有效的肌肉肿瘤局部�Objective：To investigate the anti-tumor efficacy and safety of irreversible electroporation（IRE）ablation technique in the treatment of VX_2 xenografts in muscle of rabbits in order to provide the reliable experimental evidence for the clinical application of this technique.Methods：The models of VX_2 xenografts in muscle of rabbits（n = 10） were established,then they were randomly divided into IRE-treated group（n = 8） and IRE-untreated group（as the control）（n = 2）.After treatment,the rabbits were sacrificed,and the xenograft tumors,plasma and serum samples were collected,repsectively.The tumor tissues were stained with HE.The expressions of Bcl-2,heat shock protein 70（HSP70） and vascular endothelial growth factor（VEGF） in muscular VX_2 xenograft tumors were detected by immunohistochemistry.The apoptosis of tumor cells was detected by TUNEL assay.The secretion levels of caspase 3,tumor necrosis factor-α（TNF-α） and VEGF in the plasma,the expression levels of cardiac troponinⅠ（CTnⅠ） and creatine kinase-MB isoenzyme（CKMB）,as well as the markers of liver and kidney function in serum,were detected by ELISA.Results：HE staining showed a clear borderline between the necrotic and normal areas in IRE-treated group.After IRE treatment for 3 days,there were plenty of red blood cells and infiltrated inflammatory cells in the marginal area of xenograft tumors.On the 14 th day,the fibrosis of muscle tissues appeared.IRE effectively increased the secretion levels of caspase 3 and TNF-α（both P 〈0.01）,but reduced the level of VEGF in plasma.Moreover,the Bcl-2-positive and VEGF-positive rates in tumors were decreased after IRE treatment（both P 〈0.01）,but HSP70-positive rate was diverse.The apoptosis rate of tumor cells in IREtreated groups was significantly increased as compared with the control group.After IRE treatment,the kidney function of rabbits and the levels of CTnⅠwere in the normal range,but the levels of aspartate aminotransferase（AST）,alanine transamina

关 键 词：癌 鳞状细胞 电化学疗法 肿瘤移植 细胞凋亡 免疫法 被动 

分 类 号：R73-36[医药卫生—肿瘤]