摘要:Objective To establish the pancreatic cancerogenesis model in mice and to observe the changes of the immune cell populations in peripheral blood and pancreatic lesions.Methods The in situ embedding of canerogen dimethylbenzanthracene (DMBA) was adopted to establish the pancreatic cancerogenesis process from chronic pancreatitis(CP),pancreatic intraepithelial neoplasma (PanIN) to pancreatic cancer.Total ly 60 C57BL/6J mice were used,and 8 weeks after embedding,the mice were killed.20 mice were randomized selected,and 7 immune cell populations in the peripheral blood and pancreatic lesions were detected by flow cytometery(FCM).Results During the observational peroid of 8 weeks,14(23.3%) mice died.Among the 46 survivors,20 mice were randomized selected for FCM analysis.All of the 46 pancreatic lesions were pathologically analyzed.12 cases were CP(26.1%),11 cases were lower grade PanIN (PanIN-1,2,LG-PanIN) (23.9%),9 cases were high grade PanIN (PanIN-3,HG-PanIN) (19.6%) and 14 cases were PC.Among the 20 randomized selected mice,4 cases were CP,7 cases were LG-PanIN,4 cases were HG-PanIN and 5 cases were PC.The myeloid derived suppressor cells (MDSC) of HG-PanIN and CP were significantly more than that of LG-PanIN and CP.In the pancreatic lesions,the granulocytes,MDSC and M2 polarized TAM of HG-PanIN and PC were significantly more than that of LG-PanIN and CP.On the contrary,the T lymphocytes and M1 polarized TAM were significantly decreased.Conclusions In situ embedding of DMBA is a feasible and practical method to establish the spontaneous pancreatic cancerogenesis model in the immunocompetent mice.Pancreatic cancerogenesis can induce systemic and even stronger local immunosuppression,and the MDSC and M2 polarized TAM may play the vital roles.%目的 建立小鼠胰腺癌发生模型并观察外周血及肿瘤组织中免疫细胞群的变化.方法 利用致癌剂二甲基苯蒽(DMBA)胰腺内原位包埋的方法建立从慢性胰腺炎(chronic pancreatitis,CP)、胰腺导管内瘤变(pancreatic intraepithelial neoplasma,PanIN)到胰腺癌(pancreatic cancer,PC)的发生模型.共建模60只,8周后处死小鼠.随机选择20只存活小鼠,利用流式细胞计数技术(flow cytometry,FCM)检测外周血及胰腺病变组织中7个免疫细胞群的变化.结果 建模8周内共死亡14只(23.3%),存活46只.46只小鼠病变胰腺组织均制备石蜡切片行常规HE染色.46个病变组织中,12例CP(26.1%),11例低级别PanIN (low grade-PanIN,PanIN-1,2,LG-PanIN)(23.9%),9例高级别PanIN(high grade-PanIN,PanIN-3,HG-PanIN)(19.6%)及14例PC (30.4%).随机选择的20只小鼠中,PC及HG-PanIN小鼠外周血中髓系来源抑制细胞(myeloid derived suppressor cells,MDSC)明显高于LG-PanIN及CP小鼠.PC及HG-PanIN小鼠胰腺病变组织中粒细胞、MDSC及M2型肿瘤相关巨噬细胞(tumor associated macrophages,TAM)较LG-PanIN及CP小鼠明显升高,而T淋巴细胞及M1型TAM明显减少.结论 利用DMBA胰腺内原位包埋是建立小鼠胰腺癌发生模型的简便易行的方法.胰腺癌在发生过程中,伴随全身及局部的免疫抑制效应,以病变组织内尤为明显,其中MDSC及M2型TAM可能起到关键作用.