[1]牟大鹏,王世瑶,苏冠方.抑制消减杂交技术筛选骨髓间充质干细胞与视网膜神经节样细胞差异表达基因[J].眼科新进展,2020,40(6):524-526.[doi:10.13389/j.cnki.rao.2020.0120]
 MOU Dapeng,WANG Shiyao,SU Guanfang.Selection of differentially expressed genes between bone mesenchymal stem cells and retinal ganglion-like cells by suppression subtractive hybridizationtechnique[J].Recent Advances in Ophthalmology,2020,40(6):524-526.[doi:10.13389/j.cnki.rao.2020.0120]
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抑制消减杂交技术筛选骨髓间充质干细胞与视网膜神经节样细胞差异表达基因/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年6期
页码:
524-526
栏目:
实验研究
出版日期:
2020-06-05

文章信息/Info

Title:
Selection of differentially expressed genes between bone mesenchymal stem cells and retinal ganglion-like cells by suppression subtractive hybridizationtechnique
作者:
牟大鹏王世瑶苏冠方
100730 北京市,首都医科大学附属北京同仁医院,北京同仁眼科中心,北京市眼科学与视觉科学重点实验室(牟大鹏);130041 吉林省长春市,吉林大学(王世瑶);130022 吉林省长春市,吉林大学第二医院眼科(苏冠方)
Author(s):
MOU Dapeng1 WANG Shiyao2SU Guanfang3
1.Beijing Tongren Eye Center,Beijing Tongren Hospital of Capital Medical University,Beijing Key Laboratory of Ophthalmology and Visual Sciences,Beijing 100730,China
2.Jilin University,Changchun 130041,Jilin Province,China
3.The Second Hospital of Jilin University,Changchun 130022,Jilin Province,China
关键词:
骨髓间充质干细胞抑制消减杂交视网膜神经节样细胞
Keywords:
bone marrow-derived mesenchymal stem cells suppression subtractive hybridization retinal ganglion-like cells
分类号:
R774.1
DOI:
10.13389/j.cnki.rao.2020.0120
文献标志码:
A
摘要:
目的 抑制消减杂交技术筛选和克隆视网膜神经节(RGC)样细胞与骨髓间充质干细胞的差异表达基因。方法 提取RGC样细胞总RNA为检测子,骨髓间充质干细胞的总RNA为驱动子,进而用抑制消减杂交技术方法获得消减杂交产物。结果 本消减文库共挑取克隆120个,经聚合酶链式反应(PCR)鉴定,其中含有插入片段的阳性克隆有20个。在这些阳性克隆中,PCR扩增片段大小分布于250~1500 bp。共得到5个差异基因片段,这些差异表达基因与视觉信号转导、神经细胞再生等功能有关。结论 抑制消减杂交技术有效地克隆了骨髓间充质干细胞诱导前后差异表达的基因。
Abstract:
Objective To screen and clone differentially expressed genes between retinal ganglion-like cells (RGCs) and bone marrow mesenchymal stem cells by suppression subtractive hybridization (SSH) technique. Methods The total RNA of RGCs were extracted as the detector, and the total RNA of bone marrow mesenchymal stem cells were retrieved and taken as the driver. SSH technique was used to collect subtractive hybridization products. Results A total of 120 clones were selected from the subtractive library and identified using polymerase chain reaction (PCR). Among them, 20 positive clones containing insertion fragments were identified. Among these positive clones, the amplified fragments of PCR were distributed in the range of 250 bp to 1 500 bp. There were a total of five differentially expressed genes, and they were related to visual signal transduction, neuronal regeneration, etc. Conclusion SSH technique can effectively clone differentially expressed genes of bone marrow mesenchymal stem cells before and after induction.

参考文献/References:

[1] GUAN M,XU Y,WANG W,SHAN L.Differentiation into neurons of rat bone marrow-derived mesenchymal stem cells[J].Eur Cytokine Netw,2014,25(3):58-63.
[2] 徐春玲,苏冠方,牟大鹏.乳鼠视网膜细胞诱导骨髓间充质干细胞分化为视网膜神经节样细胞[J].中华实验眼科杂志,2008,26(3):174-178.
XU C L,SU G F,MOU D P.Retinal cells of rat neonates inducing the bone marrow mesenchymal cells into RGC-like cells[J].Chin J Exp Ophthalmol,2008,26(3):174-178.
[3] 侯晓乐,李爱国,杨林,董玉珍.骨髓间充质干细胞移植对失神经肌肉萎缩大鼠的影响[J].新乡医学院学报,2015,32(5):391-394.
HOU X L,LI A G,YANG L,DONG Y Z.Effect of bone marrow mesenchymal stem cell transplantation on rats with denervated muscle atrophy[J].J Xinxiang Med Univ,2015,32(5):391-394.
[4] 滕晓坤,肖华胜.基因芯片与高通量DNA测序技术前景分析[J].中国科学(C辑),2008,38(10):891-899.
TENG X K,XIAO H S.The perspective and analysis of gene chips and high-through- put DNA sequencing approach [J].Sci Chin (Series C),2008,38(10):891-899.
[5] STRITTMATTERS M,VARTANIAN T,FISHMAN M C.GAP-43 as a plasticity protein in neuronal form and repair[J].Development Neurobiol,2010,23(5):507-520.
[6] MORGAN-LAPPE S E,TUCKER L A,HUANG X,QIAN Z ,APARNA V S,DOROTHY Z,et al.Identification of Ras-related nuclear protein,targeting protein for xenopus kinesin-like protein 2,and stearoyl-CoA desaturase 1 as promising cancer targets from an RNAi-based screen[J].Cancer Res,2007,67(9):4390-4398.
[7] KIM J,LEE S,KANG S,TAE-IL J,MAN-JONG K,TAE-HOON L,et al.Regulator of G-protein signaling 4 (RGS4) controls morphine reward by glutamate Receptor activation in the nucleus accumbens of mouse brain[J].Mol Cell,2018,41(5):454-464.
[8] YAMAGUCHI M,DAIMON Y.Overexpression of regucalcin suppresses cell proliferation in cloned rat hepatoma H4-II-E cells:Involvement of intracellular signaling factors and cell cycle-related genes[J].J Cell Bio,2010,95(6):1169-1177.
[9] FERRER I,BLANCO R,CARMONA M,PUIG B,RIBERA R,REY M J,et al.Prion protein expression in senile plaques in Alzheimer’s disease [J].Acta Neuropathol,2001,101(1):49-56.
[10] 陈娟,周琼.朊蛋白在阿尔茨海默病视网膜退行性病变中的作用研究进展[J].眼科新进展,2015,35(1):81-83,97.
CHEN J,ZHOU Q.Recent advances in role of Prion protein in Alzheimer’s retinal degeneration disease[J].Rec Adv Ophthalmol,2015,35(1):81-83,97.

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更新日期/Last Update: 2020-06-05