B7-H1在BCG治疗失败后的表达及B7-H1阻断联合BCG治疗大膀胱癌模型构建

发布时间：2018-02-01 08:55

本文关键词： 膀胱肿瘤 BCG TLR-4 B7-H1阻断　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨TLR-4和B7-H1在非肌层浸润性膀胱癌(non muscle-invasive bladder cancer,NMIBC)卡介苗(Bacillus Calmette-Guérin,BCG)免疫治疗失败患者中的表达及临床意义;构建大鼠膀胱肿瘤模型,设置B7-H1阻断与BCG联合治疗组,BCG组,B7-H1阻断组及空白对照组,实验结束后通过比较各组膀胱肿瘤的直径及B7H1表达情况以评价各组治疗方式的治疗效果,以期证实阻断B7-H1/PD-1通路能够增强BCG治疗高危膀胱癌的免疫效应。方法:选取高危NMIBC术后BCG灌注治疗失败患者20例,采用免疫组化染色法检测患者初发时和BCG治疗失败后膀胱癌组织中TLR-4和B7-H1蛋白的表达情况;构建大鼠原位膀胱肿瘤动物模型,随机分为B7-H1阻断与BCG联合治疗组,BCG组,B7-H1阻断组及空白对照组,各组处理完毕将各组大鼠处死解剖,观察大鼠膀胱及膀胱内肿瘤情况,测量各组肿瘤直径,检测各组膀胱癌细胞表面B7-H1分子表达情况。结果:BCG免疫治疗高危NMIBC患者失败后膀胱癌组织中TLR-4和B7-H1蛋白的表达较治疗前均明显增加,且二者表达呈明显正相关,差异均具有明显的统计学意义;采用膀胱内灌注N-甲基亚硝基脲(MNU)的方法,成瘤率高达95%。B7-H1阻断与BCG联合治疗组与空白对照组之间膀胱肿瘤直径差异明显,比较有统计学意义(p0.05),BCG灌注组B7-H1分子表达水平明显高于其他各组。结论:BCG免疫治疗高危NMIBC失败患者膀胱癌组织中存在着TLR-4和B7-H1表达上调,而膀胱癌BCG免疫治疗失败可能与B7-H1表达上调介导的免疫逃逸机制有关;BCG可诱导大鼠膀胱癌细胞B7-H1分子表达明显升高。阻断B7-H1/PD-1通路能增强BCG对膀胱肿瘤的治疗效果。
[Abstract]:Objective: to investigate the role of TLR-4 and B7-H1 in non muscle-invasive bladder cancer of non-myometrial invasive bladder cancer. Expression and clinical significance of NMIBC- Bacillus Calmette-Gu 茅 rinn BCG in patients with failed immunotherapy; To establish the model of bladder tumor in rats, B7-H1 block and BCG combined treatment group were set up, the BCG-H1 blocking group and the blank control group were set up. After the experiment, we compared the diameter of bladder tumor and the expression of B7H1 in order to evaluate the therapeutic effect of each group. To prove that blocking B7-H1 / PD-1 pathway can enhance the immune effect of BCG in the treatment of high risk bladder cancer. Methods: 20 patients with failed BCG perfusion therapy after high-risk NMIBC were selected. Immunohistochemical staining was used to detect the expression of TLR-4 and B7-H1 protein in bladder cancer tissues at the beginning and after the failure of BCG treatment. The animal model of in situ bladder tumor in rats was established and divided into two groups randomly: B7-H1 block group and BCG combined treatment group (BCG-H1 blocking group and blank control group). The rats in each group were sacrificed and dissected. The bladder and intravesical tumor were observed and the tumor diameter was measured. The expression of B7-H1 on bladder cancer cells was detected. The expression of TLR-4 and B7-H1 protein in bladder cancer tissues were significantly increased after the failure of BCG immunotherapy in high-risk NMIBC patients. There was a significant positive correlation between them, and the difference was statistically significant. By intravesical instillation of N-methyl-nitroso (MNU-MNU), the tumorigenesis rate was as high as 95. B7-H1 block and BCG combined treatment group and blank control group, there was significant difference in bladder tumor diameter between the two groups. The comparison was statistically significant (P 0.05). The expression of B7-H1 in BCG perfusion group was significantly higher than that in other groups. The expression of TLR-4 and B7-H1 were up-regulated in bladder cancer tissues of patients with high risk NMIBC failure of BCG immunotherapy. However, the failure of BCG immunotherapy in bladder cancer may be related to the mechanism of immune escape mediated by up-regulation of B7-H1 expression. BCG could induce the increase of B7-H1 molecule expression in bladder cancer cells, and blocking B7-H1 / PD-1 pathway could enhance the therapeutic effect of BCG on bladder cancer.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.14;R-332

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