异种MMP-2 DNA疫苗对小鼠结肠腺癌细胞CT26生长及转移的实验研究  

Active immunogene with vaccine based on chicken homologous matrix metalloproteinase-2 therapy for murine colon adencarcinoma(CT26)

作　　者：陈兢[1] 王伟[1] 游潮[1] 魏于全[2] 李浩[1] 朱彬[3] 李鹏[1] 李杨[1] 

机构地区：[1]四川大学华西医院神经外科,四川成都610041 [2]四川大学华西医院人类疾病生物治疗国家重点实验室,四川成都610041 [3]四川大学华西基础医学与法医学院,四川成都610041

出　　处：《西部医学》2008年第1期20-23,共4页Medical Journal of West China

基　　金：四川省科技厅科技攻关项目(No.04SG022-009)

摘　　要：目的通过对荷瘤裸小鼠进行异种MMP-2 DNA疫苗治疗结肠腺癌,观察其治疗效果,并初步探讨其作用机制。方法制备纯化异种MMP-2 DNA疫苗,裸鼠腋下注入大鼠CT26结肠腺癌细胞株1×106/只,建立BALB/C裸小鼠结肠腺癌动物模型,待肿瘤长至约1 cm(接种后第16天)开始治疗。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(1 mg/ml),100μl/次,每周一次,连续4周。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;Tunel法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疫苗治疗结肠腺癌能明显抑制肿瘤组织的生长,并延长荷瘤小鼠的生存期,免疫组化显示治疗组小鼠的肿瘤组织中微血管密度明显减少;Tunel法显示凋亡指数明显增多;HE染色观察肿瘤组织坏死明显增多。结论异种MMP-2 DNA疫苗治疗能够抑制肿瘤生长、抑制肿瘤血管生成,是一种独特的抗肿瘤治疗途径。Objective To evaluate whether sustained expression of chicken MMP-2 can enhance the treatment efficacy of colon adencarcinoma. Methods Constructed a c-MMP-2 DNA vaccine plasmids for DNA vaccination were purified by using two rounds of passage over Endo-free columns. CT26 tumor model was established. Briefly, CT26 cells were prepared as a slngle-cell suspension in sterile PBS at concentration of 5×10^9/1 and volume of 100 μl （1×10^6/ 200 μl cells, viability 97%）was injected i.m. into armpit of 5-to 6- weeks of age female BALB/C nude mice. When volume of tumor growth 1 cm, c-MMP-2 100 μl （1mg/ml） i.m. once weekly for 4 weeks beginning from the next day after tumor cell of inoculation. Tumor size was recorded. After the completion of therapy, microvessel density （MVD） was evaluated by immunohistochemical means （CD31） and tumor apoptosis was evaluated by terminal deoxynucleotidy Ⅰ transfer-（TdT）-mediated dUTP nlck-end labeling （TUNEL）. Results As compared with two other control groups, the treatment group with c-MMP -2 gene therapy significantly reduced tumor volume and prolonged the life span of tumor burden mice as well. In addition, the average tumor weight was lower in c-MMP -2 group than any control groups. Immunohistochemical analysis of CT26 tumors demonstrated a decreased MVD and a higher apoptosis cell rate in c-MMP -2 group. Conclusion The treatment of c-MMP-2 has an additive effect of antimetastasis and antiangiogenesis. The observation may provide an important strategy for treatment cancer metastasis.

关 键 词：异种MMP-2 DNA疫苗 结肠腺癌 转移 凋亡 

分 类 号：R730.5[医药卫生—肿瘤]