睡美人转座酶对小鼠肝内胆管癌动物模型成瘤率的影响  

Effects of sleeping beauty transposase on the tumorigenesis rate of intrahepatic cholangiocarcinoma mouse model

作　　者：周万飞 黄银久[2] 秦中强[1] 谈燚[1] ZHOU Wanfei;HUANG Yinjiu;QIN Zhongqiang;TAN Yi(Department of Hepatobiliary Surgery,the First Affiliated Hospital of Bengbu Medical College,Bengbu,Anhui 233000,China;Department of Bioscience,Bengbu Medical College,Bengbu,Anhui 233000,China)

机构地区：[1]蚌埠医学院第一附属医院肝胆外科,安徽蚌埠233000 [2]蚌埠医学院生物科学系,安徽蚌埠233000

出　　处：《重庆医学》2019年第2期297-300,316,共5页Chongqing medicine

基　　金：安徽省教育厅重大项目(KJ2018ZD022)

摘　　要：目的利用水流动力学质粒转染方法建立小鼠肝内胆管癌(ICC)动物模型,比较两种睡美人(SB)转座酶SB100与SB13对ICC动物模型成瘤率的影响。方法利用水流动力学注射方法将表达myr-Akt基因的转座子,NICD1转座子分别与SB100(A组)、SB13(B组)质粒溶液通过小鼠尾静脉注射入肝脏。注射NICD1和myr-Akt质粒作为C组,注射等量生理盐水作为D组。注射4周后处死,行病理学检查。免疫组织化学检测CK19和HA作为胆管细胞性肝癌诊断指标。结果 A组小鼠成癌率100%(20/20),B组小鼠成癌率70%(14/20),病理结果及免疫组织化学表明小鼠形成肿瘤为胆管细胞性肝癌,其中A组CK19强阳性表达率为55%,HA强阳性表达率为60%;B组CK19强阳性表达率为60%,HA强阳性表达率为65%;C、D组均无阳性表达。结论 SB100较SB13可显著提高成瘤率。Objective To establish animal model of intrahepatic cholangiocarcinoma(ICC)by hydrodynamic plasmid transfection,and to compare the effects of two transposable enzymes SB100 and SB13 on the tumorigenesis rate of intrahepatic cholangiocarcinoma animal models.Methods The transposons expressing myr-Akt gene and the NICD1 transposon were injected with the SB100(group A)and SB13(group B)transposase plasmid solution,respectively into the liver through the tail vein by hydrodynamic injection.Mice NICD1 and myr-Akt gene plasmids were injected as group C,injected with normal saline as control group D.Mice were sacrificed after 4 weeks,and cholangiocarcinoma tumor formation was determined by pathological examination and immunohistochemical detection of CK19 and HA-tag(the sequence tag of hemagglutinin surface antigen determinant of Influenza Virus).Results Group A had a cancer rate of 100%(20/20);group B had a cancer rate of 70%(14/20).Pathological results and immunohistochemistry indicated that the tumor was cholangio cellular carcinoma in group A,the strongly-positive expression rate of CK19 was 55%,the strongly-positive expression rate of HA was 60%,which in group B was 60%and 65%respectively.No positive expression in group C and D.Conclusion The transposase SB100 can significantly increase the tumor formation rate comparing with SB13.

关 键 词：模型 动物 胆管肿瘤 水流动力学质粒转染方法 转座酶类 

分 类 号：R735.8[医药卫生—肿瘤]