低剂量DEN高效诱导幼龄小鼠原发肝癌的方法  

Method of Constructing Mouse Primary Liver Cancer Model in Young Mice with Low-dose DEN

作　　者：袁玥 高玉玖 郭兴荣[1] 马石楠[1] YUAN Yue;GAO Yu-jiu;GUO Xing-rong;MA Shi-nan(Hubei Provincial Key Laboratory of Embryonic Stem Cell Research,Taihe Hospital,Hubei University of Medicine,Shiyan,Hubei 442000,China;School of Pharmacy,Hubei University of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院附属太和医院胚胎干细胞研究湖北省重点实验室 [2]湖北医药学院药学院,湖北十堰442000

出　　处：《湖北医药学院学报》2021年第4期354-358,I0001,共6页Journal of Hubei University of Medicine

基　　金：国家自然科学基金面上项目(82073232);湖北省自然科学基金面上项目(2020CFB558);湖北省教育厅科技项目重点项目(D20202103);湖北医院学院研究生创新项目(YC2020239)。

摘　　要：目的:比较不同剂量二乙基亚硝胺(DEN)和不同周龄的C57BL/6小鼠构建原发性肝癌模型的方法。方法:⑴将30只2周龄C57BL/6雄性小鼠随机分为低剂量DEN组(50 mg/kg)、高剂量DEN组(100 mg/kg),腹腔注射DEN,两周后统计小鼠死亡率;⑵将10只8周龄C57BL/6雄性小鼠随机分为PBS组和低剂量DEN组(50 mg/kg),每隔4周腹腔注射DEN一次,共8次。在小鼠44周龄时,麻醉后断颈处死取材观察肝脏外观;⑶将20只2周龄C57BL/6雄性小鼠随机分为PBS组和低剂量DEN组(50 mg/kg),每隔4周腹腔注射DEN一次,共8次,在小鼠36周龄时,每组各取5只,麻醉后断颈处死观察肝脏外观及肝组织病理学变化;剩余10只,继续观察到小鼠40周龄时,利用小动物超声仪观察肝脏成瘤情况,麻醉后断颈处死观察肝脏外观及肝组织病理学变化。结果:⑴2周龄高剂量DEN组小鼠死亡率为52.9%,而低剂量DEN组小鼠死亡率为0;⑵8周龄低剂量DEN组,在小鼠44周龄时未形成肿瘤;⑶2周龄低剂量DEN组,在小鼠36周龄时已发生癌前病变,在小鼠40周龄时已形成肝癌,小鼠成瘤率为100%。结论:2周龄小鼠低剂量DEN(50 mg/kg)诱导,在小鼠40周龄时成瘤率和存活率最高。该模型为研究原发性肝癌发生和发展机制奠定了基础。Objective To compare the methods of constructing primary liver cancer model with different doses of diethylnitrosamine(DEN)and C57BL/6 mice of different ages.Methods⑴Thirty two-week-old C57BL/6 male mice were randomly divided into low-dose DEN group(50 mg/kg)and high-dose DEN group(100 mg/kg),and the survival rate of mice was observed for two weeks after intraperitoneal injection of DEN;⑵Ten 8-week-old C57BL/6 male mice were randomly divided into PBS group and low-dose DEN group(50 mg/kg),and DEN was injected intraperitoneally every 4 weeks for a total of 8 times.When the mice were 44 weeks old,they were sacrificed after anesthesia to observe the liver appearance;⑶Twenty 2-week-old C57BL/6 male mice were randomly divided into PBS group and low-dose DEN group(50 mg/kg).DEN was injected intraperitoneally every four weeks for a total of 8 times.When the mice were 36 weeks old,five mice were taken from each group.After anesthesia,the neck was severed and sacrificed to observe the liver appearance and liver histopathological changes;The remaining ten mice were continued to be observed.At the age of 40 weeks,the liver tumorigenesis was observed by small animal ultrasound.After anesthesia,the mice were sacrificed by cervical dislocation and the liver appearance and liver histopathological changes were observed.Results⑴High-dose DEN group has a higher mortality rate of 52.9%,and low-dose DEN group(50 mg/kg)mice have a mortality rate of 0;⑵8-week-old mice did not form tumors at 44 weeks of age;⑶In the 2-week-old low-dose DEN group,precancerous lesions had occurred in the mice at 36 weeks of age,and liver cancer had formed in the mice at 40 weeks of age,and the tumor formation rate of mice was 100%.Conclusion Low-dose DEN(50 mg/kg)successfully induced liver tumors in 2-week-old mice,and the tumor formation rate and survival rate were the highest at 40 weeks old.This model lays a foundation for the study of the occurrence and development mechanism of primary liver cancer.

关 键 词：二乙基亚硝胺 原发性肝癌 小鼠 模型 

分 类 号：R735.7[医药卫生—肿瘤]