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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- 第三军医大学学报 2018年40卷第13期 页码：1171-1178
- 作者机构：
1.400014 重庆，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
2.621000 四川 绵阳，绵阳市中心医院放射科
- 作者简介：
[蔡金华，E-mail:664662256@qq.com CAI Jinhua, E-mail: 664662256@qq.com
- 基金信息：国家自然科学基金面上项目(81171387);国家自然科学基金面上项目(81771892)
DOI：10.16016/j.1000-5404.201712135
中图分类号： R329-33; R329.28; R445.2收稿：2017-12-15，
修回：2018-02-06，
纸质出版：2018-07-15
- 稿件说明：
移动端阅览
廖一凡, 蔡金华, 钟毅, 等. 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[J]. 第三军医大学学报, 2018,40(13):1171-1178.
Yifan LIAO, Jinhua CAI, Yi ZHONG, et al. Specific expression and
in vitro magnetic resonance imaging of NSE-promoter in initiating FTH1 gene during neural differentiation of bone marrow mesenchymal stem cells[J]. Journal of Third Military Medical University, 2018, 40(13): 1171-1178.廖一凡, 蔡金华, 钟毅, 等. 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[J]. 第三军医大学学报, 2018,40(13):1171-1178. DOI： 10.16016/j.1000-5404.201712135.
Yifan LIAO, Jinhua CAI, Yi ZHONG, et al. Specific expression and
in vitro magnetic resonance imaging of NSE-promoter in initiating FTH1 gene during neural differentiation of bone marrow mesenchymal stem cells[J]. Journal of Third Military Medical University, 2018, 40(13): 1171-1178. DOI： 10.16016/j.1000-5404.201712135.
摘要
目的
2
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
2
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
2
WI信号变化。
结果
2
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
2
WI信号明显降低。
结论
2
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。
Abstract
Objective
2
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
2
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 T
2
WI signal changes of both cells.
Results
2
The MSCs-NSE-FTH1 cells successfully differentiated into neuron-like cells
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 con
firmed the extensive iron accumulation in the neurons-NSE-FTH1 cells. MRI verified that the T
2
WI signal in the neurons-NSE-FTH1 cells was significantly decreased.
Conclusion
2
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
Keywords
references
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