Specific expression and <italic style="font-style: italic">in vitro</italic> magnetic resonance imaging of NSE-promoter in initiating FTH1 gene during neural differentiation of bone marrow mesenchymal stem cells

Yifan LIAO; Jinhua CAI; Yi ZHONG; Xiaoya HE; Yong QIN; Jun SUN

doi:10.16016/j.1000-5404.201712135

您当前的位置：

首页 >

文章列表页 >

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

Basic Medicine | 更新时间：2026-01-09

- Specific expression and in vitro magnetic resonance imaging of NSE-promoter in initiating FTH1 gene during neural differentiation of bone marrow mesenchymal stem cells
- Journal of Third Military Medical University Vol. 40, Issue 13, Pages: 1171-1178(2018)
- 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：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81171387);the General Program of National Natural Science Foundation of China(81771892)
- DOI：10.16016/j.1000-5404.201712135
  CLC： R329-33; R329.28; R445.2
- Received：15 December 2017，
  
  Revised：2018-02-06，
  
  Published：15 July 2018
- 稿件说明：
移动端阅览
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：

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

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

WI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

WI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。

Abstract

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 T

WI signal changes of both cells.

Results

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

WI signal in the neurons-NSE-FTH1 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

Keywords

references

DAI H Y, HE R, ZHANG Y, et al. Adenoviral vector mediated ferritin over-expression in mesenchymal stem cells detected by 7T MRI in vitro [J ] . PLoS ONE, 2017, 12(9):e0185260.DOI:10.1371/journal.pone.0185260.

STEPHEN P P, RODRIGUES T B, KETTUNEN M I, et al. Development of Timd2 as a reporter gene for MRI[J]. Magn Reson Med, 2016, 75(4):1697-1707. DOI:10.1002/mrm.25750.

PEREIRA S M, WILLIAMS S R, MURRAY P, et al. MS-1 magA: revisiting its efficacy as a reporter gene for MRI[J]. Mol Imaging, 2016, 15:1-9. DOI:10.1177/153601211 6641533.

WATADA Y, YAMASHITA D, TOYODA M, et al. Magnetic resonance monitoring of superparamagnetic iron oxide (SPIO)-labeled stem cells transplanted into the inner ear[J]. Neurosci Res, 2015, 95:21-26. DOI:10.1016/j.neures.2015.01.010.

CHO I K, MORAN S P, PAUDYAL R, et al. Longitudinal monitoring of stem cell grafts in vivo using magnetic resonance imaging with inducible maga as a genetic reporter[J ] . Theranostics, 2014, 4(10):972-989. DOI:10.7150/thno.9436.

钟毅, 蔡金华, 贺小娅, 等.骨髓间充质干细胞成神经分化中的FTH1基因表达[J].第三军医大学学报, 2016, 38(19):2121-2127.DOI:10.16016/j.1000-5404. 2016 03022.

ZHONG Y, CAI J H, HE X Y, et al. Expression of ferritin heavy chain 1 in neural differentiation of bone marrow mesenchymal stem cells[J]. J Third Mil Med Univ, 2016, 38(19):2121-2127.DOI:10.16016/j.1000-5404.2016 03022.

SONG C, WANG J, MO C, et al. Use of ferritin expression, regulated by neural cell-specific promoters in human adipose tissue-derived mesenchymal stem cells, to monitor differentiation with magnetic resonance imaging in vitro [J ] . PLoS ONE, 2015, 10(7):e0132480. DOI: 10.1371/journal.pone.0132480.

HWANG D W, KANG J J, CHUNG J, et al. Noninvasive in vivo monitoring of neuronal differentiation using reporter driven by a neuronal promoter[J ] . Eur J Nucl Med Mol Imaging, 2008, 35(1):135-145.DOI: 10.1007/s00259-007-0561-8.

贺小娅, 蔡金华, 秦勇, 等.磁共振报告基因FTH1慢病毒载体构建及其在人神经母细胞瘤细胞中的表达[J].第三军医大学学报, 2014, 36(23):2338-2342.DOI:10.16016/j.1000-5404.2014.23.002.

HE X Y, CAI J H, QIN Y, et al.Construction of recombinant lentivirus vector carrying FTH1 as an endogenous MRI reporter gene and its expression in SK-N-SH cells[J].J Third Mil Med Univ, 2014, 36(23):2338-2342. DOI:10.16016/j.1000-5404.2014.23.002.

JANOWSKI M, WALCZAK P, KROPIWNICKI T, et al. Long-term MRI cell tracking after intraventricular delivery in a patient with global cerebral ischemia and prospects for magnetic navigation of stem cells within the CSF[J]. PLoS ONE, 2014, 9(2):e97631. DOI: 10.1371/journal.pone.0097631.

DASH R, KIM P J, MATSUURA Y, et al. Manganese-enhanced magnetic resonance imaging enables in vivo confirmation of peri-infarct restoration following stem cell therapy in a porcine ischemia-reperfusion model[J ] . J Am Heart Assoc, 2015, 4(7):e002044. DOI: 10.1161/JAHA.115.002044.

NGEN E J, WANG L, KATO Y, et al. Imaging transplanted stem cells in real time using an MRI dual-contrast method[J].Sci Rep, 2015, 5:13628. DOI:10.1038/srep 13628.

HE X, CAI J, LI H, et al. In vivo magnetic resonance imaging of xenografted 3 tumors using FTH1 reporter gene expression controlled by a tet-on switch[J ] . Oncotarget, 20 16, 7(48):78591. DOI: 10.18632/oncotarget.12519.

PEREIRA S M, MOSS D, WILLIAMS S R, et al. Over-expression of the MRI reporter genes ferritin and transferrin receptor affect iron homeostasis and produce limited contrast in mesenchymal stem cells[J]. Int J Mol Sci, 2015, 16(7):15481-15496. DOI: 10.3390/ijms160715481.

GAUDET J M, RIBOT E J, CHEN Y, et al. Tracking the fate of stem cell implants with fluorine-19 MRI[J]. PLoS ONE, 2015, 10(3):e0118544. DOI:10.1371/journal.pone.0118544.

SHINOHARA Y, OHTANI T, KONNO A, et al. Viral vector-based evaluation of regulatory regions in the neuron-specific enolase (NSE) promoter in mouse cerebellum in vivo [J ] . Cerebellum, 2017, 16(5/6):913-922. DOI:10.1007/s12311-017-0866-5.

KLEIN R L, HAMBY M E, GONG Y, et al. Dose and promoter effects of adeno-associated viral vector for green fluorescent protein expression in the rat brain[J]. Exp Neurol, 2002, 176(1):66-74. DOI:10.1006/exnr.2002.7942.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Specific expression of reporter gene FTH1 driven by progression elevated gene-3 promoter in malignant transformation of rat mesenchymal stem cells and its detection by in vitro magnetic resonance imaging

Differential diagnostic efficacy of tumor vascular habitats based on DSC-PWI between glioblastoma and solitary metastatic brain tumors

Quantitative assessment of early lumbar disc degeneration in patients with low back pain by T1rho magnetic resonance imaging

Value of DCE-MRI parameters for short-term prognosis of pharyngeal cancer after postoperative radiotherapy

Application and progress of stem cells in male infertility

Related Author

孙君

蔡金华

秦勇

李晓朦

仇嘉文

LI Xuan

WANG Hanwei

ZENG Linlan

Related Institution

Department of Radiology

Chongqing Clinical Research Centre of Imaging and Nuclear Medicine

Department of Pathology， Daping Hospital， Army Medical University

Department of Radiology， the First Affiliated Hospital of Chongqing Medical University

Department of Radiology，First Affiliated Hospital， Army Medical University （Third Military Medical University）

⁰

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

DOI：10.16016/j.1000-5404.201712135

摘要

Abstract

关键词

Keywords

references