二乙基亚硝胺对肝脏过表达miR-155转基因小鼠肝癌发生的影响
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摘要
目的建立诱发性肝癌模型,观察miR-155转基因小鼠肝癌发生情况及相关基因表达变化。方法制备Rm155LG转基因小鼠,并行可视化筛选和PCR验证。利用小动物活体成像仪筛选Rm155LG/Cre双转基因小鼠,并通过荧光定量PCR(qRT-PCR)检测miR-155转基因在Rm155LG/Cre双转基因小鼠肝脏中的表达。分别对Rm155LG/Cre双转基因小鼠和同窝对照小鼠腹腔注射二乙基亚硝胺(DEN),6个月后取材行肝脏大体形态观察,并通过Western-blot分析肝癌相关基因的表达变化。结果 Luc和miR-155转基因在Rm155LG/Cre双转基因小鼠的肝脏上成功被激活。腹腔注射DEN后,与同窝对照小鼠相比,Rm155LG/Cre双转基因小鼠肝脏癌结节增多,肝癌相关基因表达上调。结论在Rm155LG/Cre双转基因小鼠上实现了miR-155转基因的肝脏特异激活,并且利用Rm155LG/Cre双转基因小鼠,初步证明了miR-155对DEN诱导的小鼠肝癌的发生有促进作用,这为后续研究miR-155在肝癌发生发展和转移中的作用奠定了坚实的基础。
Objective To observe the occurrence of hepatocellular carcinoma(HCC)in liver-specific overexpression miR-155 transgenic mice(Rm155 LG/Cre)induced by diethylnitrosamine(DEN). Methods Luc assay was used to determine Rm155 LG/Cre mice. Development of HCC was detected by morphology observation and Western blot. Results Rm155 LG/Cre mice were successfully generated. Rm155 LG/Cre mice showed accelerated growth of HCC induced by DEN and the expression of HCC related genes increased in the Rm155 LG/Cre mouse liver. Conclusions miR-155 plays an active role in the development of HCC induced by DEN. It lays a solid foundation for subsequent research on the role of miR-155 in the development and metastasis of hepatocellular carcinoma
Objective To observe the occurrence of hepatocellular carcinoma(HCC)in liver-specific overexpression miR-155 transgenic mice(Rm155 LG/Cre)induced by diethylnitrosamine(DEN). Methods Luc assay was used to determine Rm155 LG/Cre mice. Development of HCC was detected by morphology observation and Western blot. Results Rm155 LG/Cre mice were successfully generated. Rm155 LG/Cre mice showed accelerated growth of HCC induced by DEN and the expression of HCC related genes increased in the Rm155 LG/Cre mouse liver. Conclusions miR-155 plays an active role in the development of HCC induced by DEN. It lays a solid foundation for subsequent research on the role of miR-155 in the development and metastasis of hepatocellular carcinoma
引文
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[2]BHASKARAN M,MOHAN M.Micro RNAs:History,biogenesis,and their evolving role in animal development and disease[J].Vet Pathol,2014,51(4):759-774.
[3]VANNINI I,FANINI F,FABBRI M.Emerging roles of micro R-NAs in cancer[J].Curr Opin Genet Dev,2018,48:128-133.
[4]HIGGS G,SLACK F.The multiple roles of micro RNA-155 in oncogenesis[J].J Clin Bioinforma,2013,3(1):1-8.
[5]MASHIMA R.Physiological roles of mi R-155[J].Immunology,2015,145(3):323-333.
[6]CHEN Z L,MA T T,HUANG C,et al.The pivotal role of micro RNA-155 in the control of cancer[J].J Cell Physiol,2014,229(5):545-550.
[7]SONG X J,TAN S Y,WU Z C,et al.HBV suppresses ZHX2expression to promote proliferation of HCC through mi R-155activation[J].Int J Cancer,2018.DOI:10.1002/ijc.31595.
[8]LIAO W W,ZHANG C,LIU F R,et al.Effects of mi R-155 on proliferation and apoptosis by regulating Fox O3a/BIM in liver cancer cell line HCCLM3[J].Eur Rev Med Pharmacol Sci,2018,22(5):1277-1285.
[9]FU X,WEN H Q,JING L,et al.Micro RNA-155-5p promotes hepatocellular carcinoma progression by suppressing PTENthrough the PI 3K/Akt pathway[J].Cancer Sci,2017,108(4):620-631.
[10]KINEMAN R D,DEL RIO-MORENO M,SARMENTO-CA-BRAL A.40 YEARS of IGF1:Understanding the tissue-specific roles of IGF1/IGF1R in regulating metabolism using the Cre/lox P system[J].J Mol Endocrinol,2018,61(1):T187-T198.
[11]赖舒畅,邱鸿,王肖,等.条件性胰岛β细胞DEPTOR基因敲除小鼠构建及鉴定[J].实用医学杂志,2018,34(4):552-555.
[12]SCHLACHTERMAN A,CRAFT W W,HILGENFELDT E,et al.Current and future treatments for hepatocellular carcinoma[J].World J Gastroenterol,2015,21(28):8478-8491.
[13]DHIR M,MELIN A A,DOUAIHER J,et al.A review and update of treatment options and controversies in the management of hepatocellular carcinoma[J].Ann Surg,2016,263(6):1112-1125.
[14]CHIU D K C,TSE A P W,XU I M J,et al.Hypoxia inducible factor HIF-1 promotes myeloid-derived suppressor cells accumulation through ENTPD2/CD39L1 in hepatocellular carcinoma[J].Nat Commun,2017,8(1):1-12.
[15]ZHANG L,GASKINS K,YU Z Y,et al.An in vivo mouse model of metastatic human thyroid cancer[J].Thyroid,2014,24(4):695-704.
[16]BROWN Z,HEINRICH B,GRETEN T.Mouse models of hepatocellular carcinoma:An overview and highlights for immunotherapy research[J].Nat Rev Gastroenterol Hepatol,2018,15(9):536-554.
[17]赵彤,李柏.二乙基亚硝胺在小鼠诱发性肝癌模型中的应用[J].中国比较医学杂志,2016,26(1):76-80.