异种MMP-2 DNA疫苗联合小剂量顺铂治疗肿瘤转移的实验研究  

An Experimental Research on Tumor Metastasis Treated by Chicken Homologous Matrix Metalloproteinase-2 Vaccine Combined with Low-dose Cisplatin

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

机构地区：[1]四川大学华西医院神经外科,成都610041 [2]四川大学华西基础医学与法医学院生物医学工程研究室

出　　处：《四川大学学报（医学版）》2007年第1期35-39,共5页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅科技攻关项目基金(05SG022-027)资助

摘　　要：目的通过异种基质金属蛋白酶-2(MM P-2)DNA疫苗(c-MM P-2)联合小剂量的顺铂(DDP)治疗小鼠结肠癌肺转移,了解两者的协同治疗作用。方法构建编码与人类及鼠类高度同源的鸟类MM P-2分子的重组真核表达质粒作为DNA疫苗。建立负载小鼠结肠腺癌肺转移动物模型,将模型小鼠分为生理盐水对照组(N S组)、DDP治疗组(小鼠腹腔注射DDP 100μg/次,每周1次,连续2周)、c-MM P-2 DNA疫苗治疗组(小鼠后股肌肉注射1m g/mL疫苗100μL,每周1次,连续4周)、c-MM P-2 DNA疫苗联合DDP治疗组。在接种肿瘤细胞后第28 d断颈处死小鼠,取出肺并称其质量,检查并计数肺部>3 mm的肿瘤转移灶,免疫组化法测肿瘤组织微血管密度,TUNEL法测凋亡小体,EL ISA检测小鼠血清中的抗体滴度及类型。结果联合治疗组肺部肿瘤转移灶个数(3±1.64)少于c-MM P-2组(10±3.49,P=0.041)、DDP组(45±4.07,P=0.032)及N S组(85±8.70,P=0.003),肺的质量(0.4±0.03)g少于c-MM P-2组〔(0.8±0.06)g,P=0.044〕、DDP组〔(1.0±0.16)g,P=0.032〕及N S组〔(1.6±0.22)g,P=0.003〕,肿瘤组织中微血管密度低于对照组,T une l法显示凋亡小体明显增多,EL ISA法检测发现c-MM P-2治疗组和联合治疗组小鼠血清中有针对MM P-2的自身反应性抗体。结论异种MM P-2 DNA疫苗与小剂量的顺铂联合治疗恶性肿瘤转移有协同抗肿瘤转移作用。Objective To evaluate the cooperating effect on whether the chicken homologous matrix metalloproteinase-2 （c-MMP-2） vaccine combined with low-dose cisplatin （DDP） can enhance the treatment efficacy of tumor metastasis. Methods The eukaryotic expression plasmid encoding chicken homologous MMP-2 was constructed by recombinant DNA technique. In this experiment, the lung metastasis model of murine colon adenocarcinoma （C26） was established in 6- to 8- weeks of age female BALB/c mice, and then treated with 0. 9% NaCI solution （NS）, DDP, c-MMP-2, or c-MMP-2 combined DDP. The number of tumor metastasis nodules on murine lung surface was counted and the lungs were weighed after the completion of above treatments. The tumor microvessel density （MVD） and cell apoptosis were evaluated by immunohistochemical and TUNEL methods. The titer and type of autoantibodies against MMP-2 in serum of mice were evaluated by enzyme-linked immunosorbent assay （ELISA）. Results Compared with three other control groups, the combined treatment group significantly decreased the number of tumor metastasis nodules on lung surface and markedly the weight of murine lungs. The immunohistochemical analysis of tumor demonstrated a decreased MVD and a higher apoptosis cell rate happening to the combined treatment group. Autoantibodies against MMP-2 in serum of mice were, by ELISA, found in c- MMP-2 group and c-MMP-2 combined DDP group. Conclusion The antitumor effect of DDP can be potentiated by c-MMP-2. Thus the combination of c-MMP-2 and DDP results in an additive effect on the treatment of tumor metastasis.

关 键 词：基质金属蛋白酶-2 顺铂 免疫治疗 化疗 肿瘤转移 

分 类 号：R730.5[医药卫生—肿瘤]