活体荧光成像评估Ag85A和Ag85B DNA疫苗对小鼠膀胱癌移植瘤的疗效  

In vivo fluorescence image analysis system in assessing efficacies of pVAX1-Ag85A and pVAX1-Ag85B DNA vaccines in treatment of bladder cancer cell-implanted tumors in mice

作　　者：付奎[1] 杨晓峰[1] 汪海龙[1] 杨彬[1] 王炜[2] 王晔[2] 刘晶 

机构地区：[1]山西医科大学附属第一医院泌尿外科,山西太原030001 [2]浙江大学加州国际纳米技术研究院,浙江杭州31000 [3]山西离石市人民医院泌尿外科,山西离石033000

出　　处：《中国肿瘤生物治疗杂志》2009年第6期588-594,共7页Chinese Journal of Cancer Biotherapy

基　　金：山西省科技攻关基金资助项目(No.20090321022)~~

摘　　要：目的:运用活体红色荧光成像技术及影像学特征评估Ag85A DNA疫苗和Ag85B DNA疫苗对膀胱癌的免疫治疗效果。方法:构建稳定转染香菇珊瑚红色荧光蛋白(discosomasp red fluorescent protein,DsRed)基因的小鼠膀胱癌BTT细胞(BTT-DsRed),建立BTT-DsRed细胞移植瘤小鼠模型,将建模成功的24只小鼠随机分为pVAX1-Ag85A DNA疫苗组、pVAX1-Ag85B DNA疫苗组和生理盐水治疗组,各组分别于肿瘤细胞接种后的第6天肌内注射pVAX1-Ag85A、pVAX1-Ag85B及生理盐水,然后用活体荧光成像系统检测移植瘤的生长和转移情况。结果:成功制备了稳定转染DsRed基因的BTT-DsRed细胞,BTT-DsRed接种小鼠建立可视化红色荧光膀胱癌移植瘤模型。重组质粒pVAX1-Ag85A和pVAX1-Ag85B治疗后的第2周,活体荧光成像显示pVAX1-Ag85B治疗组小鼠肿瘤荧光强度明显低于生理盐水组(P<0.05);治疗后第3周,pVAX1-Ag85A和pVAX1-Ag85B组小鼠肿瘤荧光强度都明显低于生理盐水组(P<0.01),但pVAX1-Ag85B与pVAX1-Ag85A组无明显差别(P>0.05);pVAX1-Ag85B组小鼠淋巴结转移率(25.0%)明显低于生理盐水组(87.5%,P<0.01)和pVAX1-Ag85A组(62.5%,P<0.05)。结论:应用活体红色荧光成像技术能够动态、灵敏、可视化地评估DNA疫苗对小鼠膀胱癌移植瘤的疗效;Ag85A和Ag85B DNA疫苗均具有抗肿瘤免疫疗效。Objective：To use in vivo fluorescence image analysis system for evaluating the efficacies of pVAX1-Ag85A and pVAX1-Ag85B DNA vaccines in treatment of bladder cancer cell-implanted tumors in mice. Methods： Discosomasp red fluorescent protein （DsRed） stably transfected bladder cancer BTT cell line （BTT-DsRed） was established and BTT- DsRed cell-implanted mouse model was constructed. Six days later, 24 BTT-DsRed-bearing mice were randomly divided into pVAX1-Ag85A DNA vaccine group, pVAX1-Ag85B DNA vaccine group, and saline group through injecting the pVAX1-Ag85A, pVAX1-Ag85B, and saline into the right hind limbs of mice, respectively. The growth and metastasis of implanted BTF-DsRed tumors were examined by in vivo fluorescence image analysis system. Results： BTT cell line stably transfected with DsRed （BTT-DsRed） was successfully established. Fluorescence visible mouse model was successfully established by inoculating BTF-DsRed cells into hind limbs of mice. After treatment with pVAX1-Ag85A or pVAX1-Ag85B for 2 weeks, the in vivo tumor fluorescence intensity in pVAX1-Ag85B group was significantly lower than that in the saline group （P 〈 0.05 ）. After 3 weeks, tumor fluorescence intensities in both pVAX1-Ag85A and pVAX1-Ag85B groups were significantly lower than that in the saline group （ P 〈 0.01 ）. But the efficacies of pVAX1-Ag85A and pVAX1-Ag85B groups were similar （ P 〉 0. 05）. The distant lymphatic metastasis rate in pVAX1-Ag85B group was significantly lower than those in the saline （25.0% vs 87.5% ） and pVAX1-Ag85A groups （25.0% vs 62.5% ） （P 〈 0.05）. Conclusion： In vivo fluorescence image analysis system can dynamically, sensitively and visually evaluate the anti-tumor effects of DNA vaccines against bladder cancer cell-implanted tumors. Both pVAX1-Ag85A and pVAXI-Ag85B DNA vaccines have anti- tumor effects for bladder cancers.

关 键 词：AG85A AG85B DNA疫苗 膀胱肿瘤 活体荧光成像 香菇珊瑚红色荧光蛋白 

分 类 号：R737.14[医药卫生—肿瘤] R730.54[医药卫生—临床医学]