多发性骨髓瘤患者与健康人脂肪基质细胞差异的关键基因测定及分析

Key differentially expressed genes in adipose stromal cells of multiple myeloma patients versus healthy people

目的鉴定健康人与多发性骨髓瘤(multiple myeloma,MM)患者脂肪基质细胞的基因表达差异,为MM研究提供依据。方法利用基因表达数据库(Gene Expression Omnibus,GEO)下载与多发性骨髓瘤基质细胞和正常脂肪基质细胞相关的GSE133346微阵列数据芯片,通过R语言软件对数据经过标准化、校准、log2转化、转换基因ID和补充缺失值,然后以|log fold change(FC)|>1.5以及差异值P<0.05为标准筛选数据库中经校准的表达基因。差异基因通过注释、可视化和集成在线数据库(DAVID)获得GO富集分析结果,包括生物过程(biological processes,BP)、细胞成分(cellular components,CC)、分子功能(molecular functions,MF)。蛋白功能关系网络(STRING)在线数据库对筛选出的差异基因进行分析,利用图形化显示、分析和编辑蛋白网络的软件(Cytoscape)进行拓扑学分析筛选出关键差异基因。通过癌症数据在线分析和挖掘的网站(UALCAN)分析评估相关关键差异基因的表达结果。结果24个样本(MM患者和健康者各12例)来源的脂肪基质细胞,筛选出差异基因148个,其中上调47个,下调101个。GO和KEGG富集分析结果中,BP主要富集在白细胞迁移、细胞黏附、细胞对成纤维细胞生长因子刺激的反应、细胞外基质组织。CC主要富集在细胞外外泌体、内质网腔、细胞外区域、细胞表面以及质膜。MF主要富集在胶原结合,肌动蛋白结合,血小板衍生生长因子受体结合,细胞外基质结构成分以及微管蛋白结合。KEGG通路上差异基因富集在黏着力,调节肌动蛋白细胞骨架,ECM-受体相互作用,癌症中的胆碱代谢以及PI3K-Akt信号通路。拓扑学分析得到关键差异基因30个,其中上调14个,下调16个。UALCAN分析关键差异基因,其中验证出6个差异基因与肿瘤患者的生存相关,包括CSRP1、FYN、MYLK、FSTL3、LAMB1、PRRX1。其中CSRP1、FYN和MYLK与MM发展正相关,而FSTL3、LAMB1、PRRX1与MM发展负相关。结论在对12例多发性骨髓瘤患者与12名健康人脂肪基质细胞微阵列数据芯片的分析中,筛选出了6个与MM患者脂肪基质细胞相关的关键差异基因。

Objective To identify the differentially expressed of genes in adipose stromal cells in healthy people versus patients with multiple myeloma(MM),and provide evidence for MM studies.Methods The GSE133346 microarray data chip related to multiple myeloma and adipose stromal cells were downloaded from the Gene Expression Omnibus GEO.The data was standardized,calibrated,Log2 transformed,and the gene ID and supplemental deletion were converted by R language software.Then|log fold change(FC)|>1.5 and P<0.05 were used to screen the calibrated expressed genes in the database.Differentially expressed genes were inputted into the DAVID online database to obtain GO enrichment analysis results,including biological processes(BP),cellular components(CC)and molecular functions(MF).At the same time,the KEGG pathway enrichment analysis results were also obtained from the DAVID online database.The STRING online database was used to analyze the selected differentially expressed genes,and Cytoscape was used to perform the topological analysis to identify the key differentially expressed genes.The critical prognostic genes were evaluated by UALCAN of cancer data to assess the survival by differentially expressed genes.Results A total of 148 differentially expressed genes were screened for adipose stromal cells in 12 healthy controls and 12 MM patients,of which 47 were up-regulated and 101 were down-regulated.GO and KEGG enrichment analysis results showed that BP was mainly concentrated in leukocyte migration,cell adhesion,cell response to fibroblast growth factor stimulation,and extracellular matrix tissue;CC was mainly concentrated in extracellular exosomes,endoplasmic reticulum cavity,extracellular area,cell surface and plasma membrane;while MF was mainly concentrated in collagen binding,actin binding,platelet-derived growth factor receptor binding,extracellular matrix structural components,and tubulin binding.On the KEGG pathway,differentially expressed genes were enriched in adhesion,regulating actin cytoskeleton,ECM-receptor interactions,choline metabolism in cancer,and the PI3K-Akt signaling pathway.Topological analysis showed that there were 30 key differentially expressed genes,14 were up-regulated and 16 were down-regulated.UALCAN analysis showed that 6 differentially expressed genes were related to the survival of MM patients,including CSRP1,FYN,MYLK,FSTL3,LAMB1,PRRX1.Among them,CSRP1,FYN and MYLK were positively correlated with MM progressing,while FSTL3,LAMB1,PRRX1 were negatively correlated with MM progressing.Conclusion In the analysis of the microarray data chip of adipose stromal cells in multiple myeloma or healthy people,six key differentially expressed genes are identified to be related to pognosis of MM.