经组成性活化Akt1修饰的鼠胚胎肝前体细胞体内形成肿瘤的观察  

Tumor formation of fetal hepatic progenitor cells genetically modified by constitutively activated Akt1 in vivo

作　　者：马爱辉[1] 何爱丽[1,2] 葛超[1] 刘永忠[1] 

机构地区：[1]上海交通大学肿瘤研究所癌基因和相关基因国家重点实验室,上海200032 [2]苏州大学医学部基础医学与生命科学院,苏州215123

出　　处：《肿瘤》2009年第11期1059-1064,共6页Tumor

摘　　要：目的:观察组成性活化Akt1(myr-Akt1)导入的p53-/-鼠胚胎肝前体细胞(fetal hepatic progenitor cells,FHPCs)体外培养的特点和小鼠体内肿瘤模型的建立。方法:利用磁性细胞分选系统(magnetic cell-sorting system,MACS)分离上皮细胞钙黏蛋白(E-cadherin)阳性的p53-/-小鼠FHPCs,经细胞体外培养和反转录病毒导入myr-Akt1后,接种到C57BL/6小鼠的脾脏。应用RT-PCR和免疫组织化学法分析肝细胞分化和生长相关基因mRNA和蛋白的表达。结果:导入myr-Akt1后可明显提高E-cadherin阳性p53-/-小鼠FHPCs的体外生长和克隆形成能力,并可导致p16Ink4a和p19Arf mRNA表达水平下降;将该细胞接种到小鼠脾脏后可形成肿瘤,肿瘤细胞内GSK3β呈磷酸化,β-catenin呈一定水平表达。结论:成功建立了经Akt1修饰的p53-/-鼠FHPCs体内肿瘤模型,为探讨肝癌发生的分子机制和对相关抗癌药物的研究提供了平台。Objective ： To observe the biological features of constitutively activated Aktl （ myr-Aktl ） -modified p53-/-mouse fetal hepatic progenitor cells （FHPCs） in vitro and establish the tumor model in mice. Methods： E-eadherin positive p53-/-mouse FHPCs were isolated by magnetic-activated cell sorting （MACS） system. The cultured FHPCs were transfeeted with myr-Aktl mediated by ret- rovirus for cellular transformation and expansion ex vivo. The cells with the enforced expression of myr-Aktl were transplanted into the spleen of C57BL/6 mice. The mRNA and protein expressions of the genes related to differentiation and growth of liver cells were ana- lyzed by RT-PCR and immunohistochemistry. Results：Constitutively activated Aktl significantly promoted the growth and colony forma- tion of E-cadherin-positive p53-/-mouse FHPCs ex vivo, and markedly down-regulated the expression of p16＇＇k4a and p19Ar~ mRNA. The tumor formation in vivo was observed after the myr-Aktl-modified p53-/-mouse FHPCs were inoculated in the spleen of mice. GSK313 was phosphorylated and [3-catenin was expressed at certain extent in myr-Aktl-modified E-cadherin-positive p53-/-mouse FHPCs. Con- clusion：The in vivo tumor model of myr-Aktl-modified p53-/-mouse FHPCs was successfully established, which provided a useful plat- form for elucidation of the molecular mechanism for hepatocarcinogenesis and preclinical investigation of anti-cancer agents.

关 键 词：肝肿瘤 动物实验 钙黏附素 胚胎肝前体细胞 

分 类 号：R735.7[医药卫生—肿瘤] R73-352[医药卫生—临床医学]