链亲和素标记的GM-CSF治疗膀胱癌疗效的初步观察  

Preliminary Observation of Immunotherapy for Bladder Cancer by Intravesical Immobilization of GM-CSF

作　　者：梁中锟[1] 张琳[2] 谭万龙[1] 高基民[2] 陈忠[1] 黄鑫[3] 

机构地区：[1]南方医科大学附属南方医院泌尿外科,广州510515 [2]温州医学院浙江省医学遗传学重点实验室 [3]南方医科大学生命技术学院生物治疗研究所

出　　处：《临床泌尿外科杂志》2010年第11期828-831,共4页Journal of Clinical Urology

基　　金：国家863高技术研究发展计划资助项目(编号2006AA02Z4C4);广东省自然科学基金资助项目(编号9151051501000030)

摘　　要：目的:将链亲和素标记的粒细胞巨噬细胞集落刺激因子(SA-GM CSF)原位锚定成瘤小鼠膀胱,探求该新疗法的可行性和有效性。方法:选择雌性C57BL/6小鼠50只,随机均分成5组,组1到组4建立原位表浅膀胱癌模型后,按照处理的不同又分为PBS组、GM CSF组、SA-GFP组和SA-GM-CSF组,分别给予相应治疗;6次治疗后,SA-GM-CSF组和组5(对照组)接受第二次肿瘤细胞攻击;观察所有小鼠的一般情况、肿瘤情况和生存期,并对死亡小鼠进行解剖,留取器官组织做病理切片或免疫组织化学染色。结果:SA-GM CSF组与其它各组比较,肿瘤出现的时间晚,肿瘤生长速度慢,小鼠生存期长(P_(max)=0.023,P<0.05);MB49细胞二次攻击后,SAGM-CSF组与对照组相比,生存期差异有显著统计学意义(x^2=9.551.P=0.002)。结论:SA GM-CSF原位锚定于雌性C57BL/6成瘤小鼠膀胱黏膜,有效抑制了膀胱肿瘤的生长,延长了小鼠的生存时间,并且能抵抗同源肿瘤的再次攻击,提示可能诱导了小鼠针对MB49的免疫保护作用;SA-GM-CSF原位锚定治疗比单纯细胞因子灌注法疗效更好。Objective：The orthotopic mouse model of MB49 bladder cancer was used to evaluate the feasibility and efficacy of the novel immunotherapy which was given by intravesical instillation through immobilization of streptavidin （SA）-tagged bioaetive GM-CSF on the biotinylated mucosal surface of bladder wall. Methyls：Fifty fe- male C57BL/6 mice were divided into 5 groups randomly, 10 mice each group. MB49 cells were instilled to estab- lish the orthotopic mouse model of MB9 bladder cancer for mice of group 1 to group 4. One day after tumor cells instillation, NHS-PEO4-Biotin was instilled and allowed to incubate in the bladder for 0. 5 hour, followed by intra- vesical instillation of PBS, GM-CSF, SA-GFP, or SA-GM-CSF for 1 hour, respectively. The treatment was per- formed twice a week for 3 weeks. Tumor-take rate, survival, gross hematuria, and bladder weight were deter- mined as outcome variables, and the pathologic morphology of the bladders, kidney, ureter, heart, liver, lung and spleen was also observed. After 6 times of treatment, mice of SA-GM-CSF group and group 5 were pretreated and challenged with MIM9 cells ＇instilled into the bladder and monitored for survival. Results： A significant survival di{fereuce was observed between SA.-GM-CSF-treated group and arLy other groups,Tumors appeared fate,Tumors growth slowly, longer survival in mice （Pmax= 0. 023, P〈0. 05）. MB49 cells in the second attack, SA -GM - CSF group compared with the control group, survival differences were statistically significant （X2= 9. 551,P=0. 002）. Conclusions： SA - GM - CSF in situ anchored in female C57BL/6 mice into the bladder, eIIectively inhibited the growth of bladder tumors and prolong the survival time of mice, and resistant to attack again, homologous tumor; suggesting that the induction of the mouse immune protection against MB49; SA - GM - CSF treatment in situ anchored cytokines perfusion may be an attractive therapeutic alternalive for bladder cancer.

关 键 词：膀胱肿瘤 生物素 链亲和素 免疫治疗 粒细胞-巨噬细胞集落刺激因子 

分 类 号：R737.14[医药卫生—肿瘤] Q331[医药卫生—临床医学]