异种MMP-2 DNA疫苗联合伽玛刀治疗胶质瘤实验研究  

Research on treatment of active immunogene with vaccine based on chicken homologous matrix metalloproteinases-2 combined with gamma knife surgery on gliomas

作　　者：陈兢[1] 王伟[1] 游潮[1] 蔡博文[1] 李浩[1] 朱彬[2] 

机构地区：[1]四川大学华西医院神经外科伽玛刀中心 [2]四川大学华西基础医学与法医学院,四川成都610041

出　　处：《中华神经外科疾病研究杂志》2008年第1期46-50,共5页Chinese Journal of Neurosurgical Disease Research

摘　　要：目的通过对荷瘤裸小鼠进行异种基质金属蛋白酶-2(c-MMP-2)DNA疫苗联合伽玛刀(γ-刀)治疗胶质瘤,观察二者的协同治疗作用,并对其作用机制进行初步探讨。方法制备纯化c-MMP-2 DNA疫苗,裸鼠腋下注入大鼠C6胶质瘤细胞株1×106/只,建立裸鼠胶质瘤动物模型,待肿瘤长至约1cm(接种后第16d)开始治疗。γ-刀治疗以50%的等剂量曲线覆盖,边缘剂量13Gy;异种MMP-2 DNA疫苗治疗组小鼠自荷瘤后次日开始后肢肌肉注射c-MMP-2 DNA疫苗(1mg/ml),100μl/次,每周一次,连续4w。观察瘤体大小、测量肿瘤重量;免疫组化法测肿瘤组织微血管密度;末端脱氧核苷酸转移酶介导的生物素脱氧尿嘧啶核苷酸缺口标记法(TUNEL)检测各组肿瘤细胞凋亡指数;HE染色观察重要器官组织坏死情况。结果异种MMP-2 DNA疫苗联合γ-刀治疗胶质瘤能明显抑制肿瘤组织的生长,并延长荷瘤小鼠的生存期,与其他3组比较,联合治疗组免疫组化显示小鼠的肿瘤组织中微血管密度明显减少;TUNEL显示凋亡指数明显增多。结论c-MMP-2 DNA疫苗联合γ-刀治疗能够抑制肿瘤生长、抑制肿瘤血管生成,二者的联合应用具有协同效应,是一种独特的抗肿瘤治疗途径。Objective To investigate whether the sustained expression of chicken matrix metalloproteinases-2 （c-MMP-2） combined with gamma knife surgery （GKS） could enhance the therapy efficacy on gliomas. Methods Constructed c-MMP-2 DNA vaccine plasmids for DNA vaccination were purified by using two rounds of passage over endo-frec columns. Glioma model were established. Briefly, C6 cells, which were prepared as a single-cell suspension in sterile phosphate buffered solution （PBS） at concentration of 5 ×10^9/l and volume of 100 μl ×10^6/ 200μl cells, viability 97% ） were injected i.m. into armpit of 5 to 6 w female BALB/C nude mice. When the volume of tuner grew into 1 cm, GKS performed with prescription curve of 50% and prescription dose of 13 Gy. 100 μl c- MMP-2 （1 mg/ml） i.m. was performed once weekly for 4 w on 1 st day after inoculation of tuner cells. Tumor size was recorded. After the completion of therapy, Microvessel density （MVD） were evaluated by immunohistochemtry （CD 31 ） and tumor apoptesis were evaluated by terminal deoxynucleotidy transferease-mediated biotingtaled dcoxyuridine triphoephate nick-end labehng （TUNEL）. Results As compared with three other control groups, the combined treatment group with GKS and c-MMP-2 gene significantly reduced tumor volume and prolonged the life span of tuner burden mice as well. In addition, the average tumor weight was lower in GKS combined with c-MMP- 2 group than any others control groups. Inmmunohistochemical analysis of C6 gliomas demonstrated that the significant decrease of MVD in tuner tissne and a higher apoptesis cell rate in GKS combined with c-MMP-2 group. Conclusion The antitumor effect of GKS can be realized by e-MMP-2 gene therapy. These findings suggest a promising way to overcome tumor radiotherapy resistance by antiangiogenic gene therapy.

关 键 词：异种基质金属蛋白酶-2 DNA疫苗 伽玛刀 胶质瘤 

分 类 号：R741[医药卫生—神经病学与精神病学]