NSE启动子在骨髓间充质干细胞成神经分化中启动FTH1基因特异性表达及体外磁共振成像  

Specific expression and in vitro magnetic resonance imaging of NSE-promoter in initiating FTH1 gene during neural differentiation of bone marrow mesenchymal stem cells

作　　者：廖一凡[1] 蔡金华[1] 钟毅 贺小娅 秦勇[1] 孙君 LIAO Yifan;CAI Jinhua;ZHONG Yi;HE Xiaoya;QIN Yong;SUN Jun(Department of Radiology, Key Laboratory of Child Development and Disorders Cofounded by Chongqing and Ministry of Education, Chongqing Key Laboratory of Pediatrics, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Children＇s Hospital of Chongqing Medical University, Chongqing, 400014;Department of Radiology, Mianyang Central Hospital, Mianyang, Sichuan Province, 621000, China)

机构地区：[1]重庆医科大学附属儿童医院放射科儿童发育疾病研究省部共建教育部重点实验室儿科学重庆市重点实验室重庆市儿童发育重大疾病诊治与预防国际科技合作基地,重庆400014 [2]绵阳市中心医院放射科,四川绵阳621000

出　　处：《第三军医大学学报》2018年第13期1171-1178,共8页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81171387,81771892)

摘　　要：目的探讨神经元特异性烯醇化酶(neuron-specifc enolase,NSE)启动子控制的铁蛋白重链(ferritin heavy chain 1,FTH1)报告基因在骨髓间充质干细胞(bone marrow mesenchymal stem cells,MSCs)成神经分化中的特异性表达及MRI显像的可行性。方法以慢病毒为载体将含NSE启动子的FTH1基因转入MSCs中,对其成神经诱导分化,Western blot检测分化前后细胞内FTH1的表达,在0.5 mmol/L枸橼酸铁铵(ferric ammonium citrate,FAC)的培养条件下,细胞内FTH1基因表达所引起的聚铁效应通过普鲁士蓝染色及透射电镜检测;磁共振成像(magnetic resonance imaging,MRI)观察细胞T_2WI信号变化。结果含NSE启动子的FTH1基因的MSCs(MSCs-NSE-FTH1)成功分化为神经元样细胞(Neurons-NSE-FTH1)。Western blot检测结果显示FTH1在Neurons-NSE-FTH1细胞内明显表达,而在MSCs-NSE-FTH1细胞内表达极低。普鲁士蓝染色和透射电镜观察证实Neurons-NSE-FTH1细胞内较多铁颗粒聚集。MRI检查发现Neurons-NSE-FTH1细胞T_2WI信号明显降低。结论 NSE启动子在MSCs成神经分化中能特异性启动FTH1基因表达,并引起MRI信号改变。Objective To investigate the specific expression of reporter gene ferritin heavy chain 1 （FTH1） triggered by neuron-specific enolase （NSE） promoter in neural differentiation of bone marrow mesenchymal stem cells （ MSCs ）, and investigate the feasibility of its application for magnetic resonance imaging （MRI）. Methods The reporter gene FTH1 containing NSE promoter was transfected into the MSCs by using a lentivirus system. Then the genetically modified MSCs （MSCs-NSE-FTH1） were induced to differentiate into neurons （neurons-NSE-FTH1）. The dynamic expression of FTH1 in MSCs, MSCs-NSE- FTH1 and neurons-NSE-FTH1 were assessed by Western blotting. Under a cultural medium containing 0.5 mmoL/L ferric ammonium citrate, Prussian blue staining and transmission electron microscopy were used to determine the iron accumulation in the cells. MRI was then performed to detect the T2 WI signal changes of both ceils. Results The MSCs-NSE-FTH1 cells successfully differentiated into neuron-like ceils, neurons-NSE-FTH1. Western blotting showed that FTH1 was expressed significantly in the neurons-NSE-FTH1 cells, but mildly in MSCs-NSE-FTH1 cells. Prussian blue staining and transmission electron microscopy confirmed the extensive iron accumulation in the neurons-NSE-FTHl ceils. MRI verified that the T2WI signal in the neurons-NSE-FFH1 cells was significantly decreased. Conclusion The NSE promoter could specifically initiate the expression of reporter gene FTH1, and then result in an obvious MRI signal change, during the differentiation of MSCs into neurons.

关 键 词：NSE启动子 FTH1报告基因 磁共振成像 干细胞 神经分化 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]