Vang Daniel P, Wurz Gregory T, Griffey Stephen M, Kao Chiao-Jung, Gutierrez Audrey M, Hanson Gregory K, Wolf Michael, DeGregorio Michael W
Department of Internal Medicine, Division of Hematology and Oncology, University of California, Davis.
J Vis Exp. 2013 Oct 30(80):e50868. doi: 10.3791/50868.
A preclinical model of invasive bladder cancer was developed in human mucin 1 (MUC1) transgenic (MUC1.Tg) mice for the purpose of evaluating immunotherapy and/or cytotoxic chemotherapy. To induce bladder cancer, C57BL/6 mice (MUC1.Tg and wild type) were treated orally with the carcinogen N-butyl-N-(4-hydroxybutyl)nitrosamine (OH-BBN) at 3.0 mg/day, 5 days/week for 12 weeks. To assess the effects of OH-BBN on serum cytokine profile during tumor development, whole blood was collected via submandibular bleeds prior to treatment and every four weeks. In addition, a MUC1-targeted peptide vaccine and placebo were administered to groups of mice weekly for eight weeks. Multiplex fluorometric microbead immunoanalyses of serum cytokines during tumor development and following vaccination were performed. At termination, interferon gamma (IFN-γ)/interleukin-4 (IL-4) ELISpot analysis for MUC1 specific T-cell immune response and histopathological evaluations of tumor type and grade were performed. The results showed that: (1) the incidence of bladder cancer in both MUC1.Tg and wild type mice was 67%; (2) transitional cell carcinomas (TCC) developed at a 2:1 ratio compared to squamous cell carcinomas (SCC); (3) inflammatory cytokines increased with time during tumor development; and (4) administration of the peptide vaccine induces a Th1-polarized serum cytokine profile and a MUC1 specific T-cell response. All tumors in MUC1.Tg mice were positive for MUC1 expression, and half of all tumors in MUC1.Tg and wild type mice were invasive. In conclusion, using a team approach through the coordination of the efforts of pharmacologists, immunologists, pathologists and molecular biologists, we have developed an immune intact transgenic mouse model of bladder cancer that expresses hMUC1.
为了评估免疫疗法和/或细胞毒性化疗,在人黏蛋白1(MUC1)转基因(MUC1.Tg)小鼠中建立了浸润性膀胱癌的临床前模型。为诱导膀胱癌,C57BL/6小鼠(MUC1.Tg和野生型)口服给予致癌物N-丁基-N-(4-羟丁基)亚硝胺(OH-BBN),剂量为3.0mg/天,每周5天,共12周。为评估OH-BBN在肿瘤发生过程中对血清细胞因子谱的影响,在治疗前及每四周通过下颌下出血采集全血。此外,将MUC1靶向肽疫苗和安慰剂每周给予小鼠组,共八周。在肿瘤发生过程中和接种疫苗后,对血清细胞因子进行多重荧光微珠免疫分析。在实验结束时,进行干扰素γ(IFN-γ)/白细胞介素-4(IL-4)ELISpot分析以检测MUC1特异性T细胞免疫反应,并对肿瘤类型和分级进行组织病理学评估。结果显示:(1)MUC1.Tg和野生型小鼠的膀胱癌发生率均为67%;(2)移行细胞癌(TCC)与鳞状细胞癌(SCC)的发生比例为2:1;(3)在肿瘤发生过程中,炎性细胞因子随时间增加;(4)给予肽疫苗可诱导Th1极化的血清细胞因子谱和MUC1特异性T细胞反应。MUC1.Tg小鼠中的所有肿瘤MUC1表达均为阳性,MUC1.Tg和野生型小鼠中所有肿瘤的一半为浸润性。总之,通过药理学家、免疫学家、病理学家和分子生物学家的共同努力,我们开发了一种表达hMUC1的免疫健全的膀胱癌转基因小鼠模型。