SIRT6在二乙基亚硝胺诱导小鼠自发肝癌中的作用  

Effect of SIRT6 on spontaneous hepatocellular carcinoma induced by diethyl nitrosamine in mice

作　　者：朱会生 张翠翠[1] 范蕊[1] 熊熙文 千新来[1] ZHU Huisheng;ZHANG Cuicui;FAN Rui;XIONG Xiwen;QIAN Xinlai(Department of Pathology,Basic Medical College,Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Department of General Surgery,Chinese People′s Liberation Army 83 Group Army Hospital,Xinxiang 453000,Henan Province,China;Laboratory of Molecular Metabolism,School of Forensic Medicine,Xinxiang Medical University,Xinxiang 453003,Henan Province,China)

机构地区：[1]新乡医学院基础医学院病理学教研室,河南新乡453003 [2]中国人民解放军陆军第八十三集团军医院普通外科,河南新乡453000 [3]新乡医学院法医学院分子代谢实验室,河南新乡453003

出　　处：《新乡医学院学报》2020年第4期332-336,共5页Journal of Xinxiang Medical University

基　　金：国家自然科学基金面上项目(编号:81670526);河南省科技攻关项目(编号:182102310107);河南省基础与前沿技术项目(编号:122300410121);新乡市科技攻关项目(编号:CXGG17007)。

摘　　要：目的探讨SIRT6在二乙基亚硝胺(DEN)诱导小鼠自发肝癌中的作用。方法选取Huh7-sirt6细胞和Huh7-GFP细胞,常规培养,细胞计数试剂盒(CCK-8)法检测过表达SIRT6对肝癌细胞体外增殖的影响。选取Wt小鼠和TgSIRT6小鼠各1只,Western blot法检测TgSIRT6小鼠肝脏组织中SIRT6表达;另选取15日龄清洁级无病原体全身性过表达SIRT6转基因(TgSIRT6)小鼠20只(雄性14只,雌性6只)和C57BL6/J野生型(Wt)小鼠18只(雄性12只,雌性6只)为研究对象,将小鼠分为TgSIRT6-DEN组16只(雄性10只,雌性6只)、Wt-DEN组14只(雄性8只,雌性6只)、TgSIRT6对照组4只(雄性)和Wt对照组4只(雄性)。TgSIRT6-DEN组和Wt-DEN组小鼠腹腔注射DEN25 mg·kg-1;TgSIRT6对照组和Wt对照组小鼠腹腔注射相同剂量的生理盐水。给药后48周颈椎脱臼法处死各组小鼠,取肝脏,观察肝脏颜色、大小、质地及有无癌变结节,记录肝脏肿瘤的大小、数量,并行肝肿瘤组织苏木精-伊红染色。结果细胞培养第4天,Huh-SIRT6细胞增殖速度显著低于Huh-GFP细胞(P<0.05)。Wt小鼠和TgSIRT6小鼠肝脏组织中SIRT6的表达量分别为0.247±0.055和1.178±0.037,TgSIRT6小鼠肝脏组织中SIRT6的表达量高于Wt小鼠(P<0.05)。TgSIRT6对照组和Wt对照组小鼠肝脏中未观察到肿瘤;TgSIRT6-DEN组雄鼠和雌鼠的致瘤率分别为80%(8/10)和33%(2/6),Wt-DEN组雄鼠和雌鼠的致瘤率分别为100%(8/8)和83%(5/6);TgSIRT6-DEN组雄鼠和雌鼠肿瘤数量及最大肿瘤直径均小于Wt-DEN组(P<0.05)。HE染色结果显示,肿瘤为肝细胞癌。结论肝细胞中SIRT6过表达能抑制DEN诱发的肝脏肿瘤的形成。Objective To explore the effect of SIRT6 on spontaneous hepatocellular carcinoma induced by diethyl nitrosamine(DEN)in mice.Methods The Huh7-sirt6 cells and Huh7-GFP cells were cultured conventionally,cell count kit was used to detect the effect of overexpression of SIRT6 on the proliferation of hepatocellular carcinoma cells in vitro.One wild type(WT)mice and One TgSIRT6 mice were selected and the expression of h SIRT6-HA in the liver of mice was detected by Western blot.Twenty-one 15 days old mice(male 14,female 6)of transgenic gene of SIRT6 overexpression in whole body(TgSIRT6)without pathogens and 19 Wt C57 BL6/J mice(12 male mice,6 female mice)were selected as the research object.The mice were divided into TgSIRT6-DEN group(n=16,10 male mice,6 female mice),Wt-DEN group(n=14,8 male mice,6 female mice),TgSIRT6 control group(n=4,all male)and Wt control only four(n=4,all male).Mice in the tg SIRT6-DEN group and Wt-DEN group were intraperitoneally injected with 25 mg·kg-1 DEN.Mice in the TgSIRT6 control group and Wt control group were intraperitoneally injected with the same dose of normal saline.The mice was sacrificed by cervical dislocation at 48 weeks after administration,and the livers were obtained to observe the color,size,texture and presence of cancerous nodules.The size and the number of liver tumors were recorded.Hematoxylin-eosin staining was used to observe the liver tissue.Results On the 4 thday of cell culture,the proliferation rate of Huh-sirt6 cells was significantly lower than that of Huh-GFP cells(P<0.05).The expression levels of SIRT6 in the liver tissues of Wt mice and TgSIRT6 mice were 0.247±0.055 and 1.178±0.037,respectively,and the expression levels of SIRT6 in the liver tissues of TgSIRT6 mice were higher than that of Wt mice(P<0.05).No tumor was observed in the liver in the TgSIRT6 control group and Wt control group.The tumorigenic rate of male and female mice in the TgSIRT6-DEN group were 80%(8/10)and 33%(2/6),respectively.The tumorigenic rate of male and female mice in Wt-DEN group

关 键 词：二乙基亚硝胺 肝脏自发肿瘤 SIRT6 转基因小鼠 

分 类 号：R735.7[医药卫生—肿瘤]