重组卡介苗-白细胞介素-15-Ag85B的构建及其抗膀胱癌作用  

Construction of recombinant Bacillus Calmette-Guerin-interleukin-15-AG85B and its anti-bladder cancer effect

作　　者：范毛川[1] 李杨东 杨晓亮 窦启锋[1] Fan Maochuan;Li Yangdong;Yang Xiaoliang;Dou Qifeng(The First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,China;Department of Urology,Hefei Central Hospital,Hefei 230022,China)

机构地区：[1]新乡医学院第一附属医院,卫辉453100 [2]合肥市中心医院泌尿外科,230022

出　　处：《中华实验外科杂志》2022年第7期1337-1340,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划项目(LHGJ20190449);2021年科新乡市灾后重建科技专项(21CJ002)。

摘　　要：目的探讨重组卡介苗(BCG)的抗膀胱癌作用,评估膀胱内灌注重组BCG对大鼠生存的影响。方法构建稳定表达白细胞介素(IL)-15和Ag85B的重组BCG菌株(BCG-IL-15-Ag85B),建立大鼠膀胱癌模型,分为BCG-IL-15-Ag85B灌注组,BCG灌注组和磷酸盐缓冲液(PBS)灌注组。观察并记录各组大鼠的生存情况,测量各组大鼠肿瘤的体积,以评估抗肿瘤效果;流式细胞术检测不同组别膀胱组织内嗜中性粒细胞和淋巴细胞的水平,荧光定量反转录-聚合酶链反应(RT-PCR)检测不同组别中巨噬细胞炎症蛋白1α(MIP1α)和MIP2 mRNA的表达水平。组间比较采用独立样本t检验。结果成功构建可表达IL-15融合蛋白的重组BCG,BCG-IL-15-Ag85B可显著延长荷瘤大鼠的生存时间以及抑制肿瘤的生长。BCG-IL-15-Ag85B灌注组(607.33±18.45)的中性粒细胞数目明显高于BCG灌注组(405.00±24.64)和PBS灌注组(204.67±20.55),差异有统计学意义(t=11.386,P<0.05);BCG-IL-15-Ag85B灌注组(114.67±15.82)的淋巴细胞数目明显高于BCG灌注组(50.00±10.00)和PBS灌注组(17.67±2.52),差异有统计学意义(t=5.984,P<0.05)。BCG-IL-15-Ag85B灌注组(2.78±0.57)的MIP1αmRNA的表达水平明显高于BCG灌注组(1.76±0.13)和PBS灌注组(1.00±0.06),差异有统计学意义(t=5.327,P<0.05)。BCG-IL-15-Ag85B灌注组(2.96±0.62)的MIP2 mRNA的表达水平明显高于BCG灌注组(1.88±0.16)和PBS灌注组(1.00±0.07),差异有统计学意义(t=5.674,P<0.05)。结论膀胱内灌注BCG-IL-15-Ag85B增强了膀胱组织的免疫应答,提高了抗肿瘤效果,为膀胱癌的临床治疗提供新思路和新策略。Objective To investigate the anti-bladder cancer effect of recombinant Bacillus Calmette-Guerin(BCG),further to evaluate the effect of intravesical perfusion of recombinant BCG on the survival of rats.Methods The recombinant BCG strain[BCG-interleukin(IL)-15-Ag85B]stably expressing IL-15 and Ag85B was constructed,and the rat model of bladder cancer was established,which was divided into BCG-IL-15-Ag85B perfusion,BCG perfusion and phosphate buffer saline(PBS)perfusion groups.The survival of the rats in each group was observed,and the tumor volume in each group was measured.The counts of neutrophils and lymphocytes in different groups of bladder tissues were detected by flow cytometry,and the mRNA expression levels of macrophage inflammatory protein 1alpha(MIP1α)and MIP2 in different groups were detected by fluorescence quantitative reverse transcription-polymerase chain reaction(RT-PCR).Two independent samples t-test was used for comparison between groups.Results Recombinant BCG expressing IL-15 fusion protein was successfully constructed.BCG-IL-15-Ag85B could significantly prolong the survival time of tumor-bearing rats and inhibit tumor growth.The counts of neutrophils in the BCG-IL-15-Ag85B perfusion group(607.33±18.45)were significantly greater than those in the BCG perfusion group(405.00±24.64)and PBS perfusion group(204.67±20.55,t=11.386,P<0.05).The counts of lymphocytes in the BCG-IL-15-Ag85B perfusion group(114.67±15.82)were significantly greater than those in the BCG perfusion group(50.00±10.00)and PBS perfusion group(17.67±2.52,t=5.984,P<0.05).The expression level of MIP1αmRNA in BCG-IL-15-Ag85B perfusion group(2.78±0.57)was significantly higher than that in BCG perfusion group(1.76±0.13)and PBS perfusion group(1.00±0.06,t=5.327,P<0.05).The expression level of MIP2 mRNA in the BCG-IL-15-Ag85B perfusion group(2.96±0.62)was significantly higher than that in the BCG perfusion group(1.88±0.16)and PBS perfusion group(1.00±0.07,t=5.674,P<0.05).Conclusion Intravesical infusion of BCG-IL-15-Ag85B e

关 键 词：膀胱癌 重组卡介苗 免疫治疗 白细胞介素-15 

分 类 号：R737.14[医药卫生—肿瘤]