基础医学与临床 ›› 2015, Vol. 35 ›› Issue (8): 1078-1082.

• 研究论文 • 上一篇    下一篇

间充质干细胞与肝样转化细胞的基因谱差异分析

姜华1,高建鹏2,王辉3,张志波3,王丽昆3   

  1. 1. 昆明医科大学附属延安医院消化内科
    2. 昆明市延安医院消化内科
    3. 昆明医科大学附属延安医院
  • 收稿日期:2014-11-27 修回日期:2015-03-21 出版日期:2015-08-05 发布日期:2015-07-15
  • 通讯作者: 高建鹏 E-mail:gaojianpengkm@163.com

To analysis differencied gene expression between mesenchymal stem cells and hepatocyte-like transformed cells

  • Received:2014-11-27 Revised:2015-03-21 Online:2015-08-05 Published:2015-07-15

摘要: 目的 用全基因表达谱芯片筛选大鼠骨髓间充质干细胞(MSCs)与定向分化为肝细胞样细胞(HLCs)的表达基因差异并对其进行分析研究。方法 将大鼠MSCs诱导定向分化的肝样细胞做3次生物学重复,分别取MSCs和诱导分化的肝样细胞提取总RNA进行扩增、Cy3标记。与大鼠全基因表达谱芯片杂交,荧光扫描,筛选出差异表达基因,并进行GO分析和pathway分析。用实时定量PCR对结果进行验证。结果 芯片筛选结果显示,分化后的MSCs与HLCs间的差异表达的基因多达839个,其中上调表达448个, 下调表达391个。在差异基因当中有生物学过程注释的基因有363个,具有细胞组分注释的差异基因有377个,具有分子功能注释的差异基因364个。Pathwav分析结果中显示在差异基因参与的273个Pathway中有显著性变化的Pathway有45个,其中参与到显著性Pathway共有275个差异表达基因,其中上调基因122个,下调基因153个。通过差异基因构建基因调控网络,按照基因在网络的度(Degree)筛选出网络当中的5个关键基因:Pkm2、 Nt5e 、Fos、 Adcy3 、Itga7。结论 MSCs体外诱导定向分化为HLCs过程中,其调节的基因与肝细胞中细胞增殖、分化、凋亡、代谢和调控等密切相关。

关键词: 【关键词】:全基因组表达芯片, 间充质干细胞, 肝样细胞, GO分析, Pathway分析

Abstract: Objective To analysis of differential on gene expression profiles between rat bone marrow mesenchymal stem cells and hepatocyte like cells which were directional differentiated by gene expression microarray technology. Method To make the rat’s MSCs directional differentiation into hepatocyte-like cells this procedure was repeated 3 times, to amplified respectively MSCs and hepatocyte-like cell total RNA, fluorescent labeled Cy3 probe. And then basing on fluorescence in situ hybridization, scanning, screening the differential information with GO and pathway analysis. The results were validated by RT-PCR. Results The test results appeared to MSCs Vs HLCs 839 genes was different , among 448 genes up-regulated, 391 genes down –regulated,363 genes with biological process annotation, 377 genes with cellular components annotation, 364 genes with molecular function annotation. The Pathway analysis results revealed 45 pathways had significant changed among the 273 Pathways, 275 differentially expressed genes involved in those pathway, including 122 up-regulated genes and 153 down regulated genes. Constructing gene regulatory network through those differential gene expression, according to the degree in the network and selected 5 key gene: Pkm2, Nt5e, Fos, Adcy3, Itga7. Conclusion In this procedure that MSCs was induced and differentiate into HLCs in vitro, These regulate gene expression in transformed cells just as the same as liver cells, such as: proliferation, differentiation, apoptosis, metabolism and regulation are closely related.

Key words: 【Keywords】: whole genome expression chip, mesenchymal stem cells, hepatocyte like cells, GO analysis, Pathway analysis