肝脏IKK2持续性激活对Myc过表达诱导的小鼠原发性肝肿瘤发生发展的影响  

Effects of liver constitutive activation of IKK2 on the development of mouse hepatocarcinogenesis induced by Myc overexpression

作　　者：贺佳佳 韦炜[1] 丁昀 吴骏[1] 胡文蔚[1] 蒋敬庭[2] HE Jiajia;WEI Wei;DING Yun;WU Jun;HU Wenwei;JIANG Jingting(Department of Oncology,The Third Affiliated Hospital of Soochow University,Cancer Biology Diagnosis and Treatment Center of the Third Affiliated Hospital of Soochow University,Changzhou 213003,China)

机构地区：[1]苏州大学附属第三医院肿瘤科苏州大学附属第三医院肿瘤生物诊疗中心,江苏常州213003 [2]江苏省肿瘤免疫治疗工程技术研究中心苏州大学细胞治疗研究院,213003

出　　处：《临床肿瘤学杂志》2022年第8期673-679,共7页Chinese Clinical Oncology

基　　金：国家重点研发资助项目(2018YFC1313400);国家科技支撑计划资助项目(2015BAI12B12);国家自然科学基金海外及港澳学者合作研究基金项目(31729001);国家自然科学基金资助项目(81972869);江苏省重点研发计划专项资金项目(BE2018645)。

摘　　要：目的探讨持续性激活IKK/核因子(NF)-κB信号通路对Myc过表达诱导的小鼠原发性肝癌发生发展的影响。方法利用转基因小鼠将LAP-tTA小鼠与Alb-cre小鼠杂交,再与CAG-LSL-IKK2-CAki小鼠杂交以产生LAP-tTA+/Alb-cre+/CAG-LSL-IKK2-CAki小鼠,最后与tetO-Myc^(+)小鼠杂交。本实验包括4组动物:WT小鼠(对照)、IKK2-CA^(hep)小鼠(肝细胞中IKK2持续激活且无Myc过表达)、MYC^(LAP-tTA)小鼠(Myc过表达且无IKK2持续激活)和MYC^(LAP-tTA)/IKK2-CA^(hep)小鼠(肝脏Myc过表达和IKK2持续激活)。记录小鼠肝脏质量和体质量,检测肝脏酶学指标,记录肝脏大体形态,H&E染色观察肝组织病理结构,天狼星红染色检测肝脏纤维化,免疫荧光染色检测肝脏炎症指标、肝前体细胞标志物,实时荧光定量PCR检测促炎症反应基因和促纤维化反应基因的mRNA水平。结果MYC^(LAP-tTA)/IKK2-CA^(hep)小鼠对比对照组小鼠,肝脏/体质量比值升高[(29.360±1.585)%vs.(4.967±0.1909)%],谷草转氨酶[(173.7±28.99)U/L vs.(45.67±19.12)U/L]、谷丙转氨酶[(86.93±19.02)U/L vs.(22.04±10.09)U/L]和碱性磷酸酶[(1680±940.6)U/L vs.(239.1±43.32)U/L]水平均上调,差异有统计学意义(P<0.05)。MYC^(LAP-tTA)/IKK2-CA^(hep)小鼠对比MYC^(LAP-tTA)小鼠,促炎症反应基因Emr1(76.30±10.67 vs.36.20±15.64)及促纤维化反应基因Col1a1(51.67±5.613 vs.17.29±1.350)、Col3a1(61.33±13.85 vs.18.37±5.547)和Vimentin(62.20±17.16 vs.28.23±4.952)的mRNA水平均升高(P<0.05)。IKK2-CA^(hep)及MYC^(LAP-tTA)/IKK2-CA^(hep)小鼠较对照组的CD45表达显著增加,IKK2-CA^(hep)小鼠肝细胞周围有大面积的胶原纤维表达。结论肝细胞中持续性活化IKK/NF-κB经典信号通路,可促进肝脏炎性反应及肝脏纤维化并诱导肝前体细胞的生成。Objective To investigate the role of IKK/nuclear factor(NF)-κB signaling pathway in the development of Myc-induced mouse hepatocellular carcinoma and potential mechanisms.Methods Transgenic mice were studied in the project.LAP-tTA mice were crossed with Alb-cre transgenic mice,and the offsprings were crossed with CAG-LSL-IKK2-CAki mice,which lastly were crossed to tetO-Myc^(+)mice.Four groups of animals were included:WT mice(no expression of Myc and no IKK2 activetion),IKK2-CA^(hep) mice(specific activation of IKK2 in liver parenchymal cells without Myc overexpression),MYC^(LAP-tTA) mice(only Myc overexpression without activation of IKK2)and MYC^(LAP-tTA)/IKK2-CA^(hep) mice(both overexpression of Myc and activation of IKK2).The liver mass and body mass of mice were recorded.Liver enzyme levels were tested.Mouse liver was observed both macroscopically and microscopically.H&E staining was used to observe the pathology of liver tissues.Sirius red staining was performed to detect liver fibrosis.Immunofluorescence staining was performed to detect liver inflammatory marker and liver progenitor cell markers.The mRNA levels of pro-inflammatory genes and pro-fibrogenic genes were detected by real-time quantitative reverse transcriptase-polymerase chain reaction in each group.Results The liver/weight mass ratio was significantly elevated in MYC^(LAP-tTA)/IKK2-CA^(hep) mice compared to control mice[(29.36±1.585)%vs.(4.967±0.1909)%,P<0.05)].Serum levels of aspartate aminotransferase[(173.7±28.99)U/L vs.(45.67±19.12)U/L],alanine transaminase[(86.93±19.02)U/L vs.(22.04±10.09)U/L]and alkaline phosphatase[(1680±940.6)U/L vs.(239.1±43.32)U/L]were significantly increased in livers of MYC^(LAP-tTA)/IKK2-CA^(hep) mice compared to control mice(P<0.05).The mRNA levels of pro-inflammatory genes such as Emr1(76.30±10.67 vs.36.20±15.64)and pro-fibrogenic genes such as Col1a1(51.67±5.613 vs.17.29±1.350),Col3a1(61.33±13.85 vs.18.37±5.547)and Vimentin(62.20±17.16 vs.28.23±4.952)were significantly increased in MYC^(LAP-tTA)/

关 键 词：肝癌 IKK2 MYC 肝纤维化 肝前体细胞 

分 类 号：R735.7[医药卫生—肿瘤]