[1]朱会生,张翠翠,范　蕊,等.SIRT6在二乙基亚硝胺诱导小鼠自发肝癌中的作用[J].新乡医学院学报,2020,37(4):332-336.[doi:10.7683/xxyxyxb.2020.04.007]
　ZHU Huisheng,ZHANG Cuicui,FAN Rui,et al.Effect of SIRT6 on spontaneous hepatocellular carcinoma induced by diethyl nitrosamine in mice[J].Journal of Xinxiang Medical University,2020,37(4):332-336.[doi:10.7683/xxyxyxb.2020.04.007]
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SIRT6在二乙基亚硝胺诱导小鼠自发肝癌中的作用
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
37
期数:
2020年4
页码:
332-336
栏目:
基础研究
出版日期:
2020-04-05

文章信息/Info

Title:
Effect of SIRT6 on spontaneous hepatocellular carcinoma induced by diethyl nitrosamine in mice
作者:
朱会生1; 2; 张翠翠1; 范　蕊1; 熊熙文3; 千新来1
（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）
Author(s):
ZHU Huisheng1; 2; ZHANG Cuicui1; FAN Rui1; XIONG Xiwen3; QIAN Xinlai1
(1.Department of Pathology,Basic Medical College,Xinxiang Medical University，Xinxiang 453003，Henan Province，China；2.Department of General Surgery,Chinese People′s Liberation Army 83 Group Army Hospital,Xinxiang 453000，Henan Province，China;3.Laboratory of Molecular Metabolism,School of Forensic Medicine,Xinxiang Medical University，Xinxiang　453003，Henan Province，China)
关键词:
二乙基亚硝胺; 肝脏自发肿瘤; SIRT6; 转基因小鼠
Keywords:
diethyl nitrosamine; spontaneous hepatocellular carcinoma; SIRT6; transgenic mouse
分类号:
R735.7
DOI:
10.7683/xxyxyxb.2020.04.007
文献标志码:
A
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DEN 25 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。
Abstract:
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 hSIRT6-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 C57BL6/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 tgSIRT6-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 4th day 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 were 100%(8/8) and 83%(5/6).The number and maximum diameter of tumor in the tgSIRT6-DEN group in both male and female mice were smaller than those in Wt-DEN group (P<0.05).HE staining showed that the tumor was hepatocellular carcinoma.Conclusion　Overexpression of SIRT6 in hepatocytes can inhibit the formation of liver tumors induced by DEN.

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更新日期/Last Update: 2020-04-05