单独骨髓复发儿童急性B淋巴细胞白血病的生物信息学分析及验证

Bioinformatics Analysis and Verification of Acute B Lymphocytic Leukemia in Children with Isolated Bone Marrow Relapse

目的:分析影响儿童急性B淋巴细胞白血病单独骨髓复发患儿预后的枢纽基因。方法:从TCGA数据库下载符合要求的高通量RNA测序数据,运用R语言DESeq2程序包筛选出差异表达基因,并对这些基因进行GO功能富集分析和KEGG通路富集分析,同时使用STRING数据库数据和Cytoscape软件构建蛋白质相互作用网络,筛选枢纽基因和高度蛋白互作子网络并分别进行功能富集和通路富集分析,用JASPAR数据库筛选枢纽基因启动子上游转录因子,结合TCGA数据库附带的临床信息对枢纽基因进行生存分析,进而采集临床患者的骨髓样本和病历资料,通过临床样本验证枢纽基因分析结果。结果:共筛选出847个差异表达基因,其中上调的差异表达基因813个,下调的差异表达基因34个,筛选出枢纽基因11个。RPS5、RPS15、RPL23、RPL35、RPS8、RPS27A、RPS3、RPL9、RPS21、RPS7、RPL38对患儿生存有明显影响,且ZNF460可能参与它们的调控,其中RPS3、RPS15、RPS8、RPS27A、RPS21的高表达在复发患者的临床样本得到了验证。结论:RPS3、RPS15、RPS8、RPS27A、RPS21有望作为提示单独骨髓复发这一恶性事件的生物标志物,且可能被ZNF460调控。

qAbstract Objective: To analyze the hub genes affecting the solely bone marrow relapse of childish acute B-cell lymphoblastic leukemia( B-ALL). Methods: The high-throughput RNA sequencing data were downloaded from TCGA database,the differentially expressed genes were screened by DESeq2 package of R,and the differentially expressed genes were grouped by GO function enrichment analysis and KEGG pathway enrichment analysis. Further,the data of STRING database and Cytoscape software were used to construct protein interaction network,screen hub genes and highly interaction protein sub network,perform GO and KEGG analysis of the hub genes and protein sub network respectively. JASPAR database was used to screen the upstream transcription factor of the hub gene promoter. Survival analysis based on the expression of hub genes was performed with clinical information attached to TCGA database. The bone marrow samples and clinical data of the patients were collected,the analysis results of hub genes were verified through clinical samples. Results: 847 differentially expressed genes were collected,including 813 up-regulated genes,34 down-regulated genes,11 hub genes were screened out. The results of survival analysis showed that RPS5、RPS15、RPL23、RPL35、RPS8、RPS27 A、RPS3、RPL9、RPS21、RPS7 and RPL38 showed significant effect on the survival of the children,and ZNF460 might be involved in their regulation. The high expressions of RPS3,RPS15,RPS8,RPS27 A,and RPS21 had been verified in clinical samples of solely bone marrow relapsed patients. Conclusion: RPS3,RPS15,RPS8,RPS27 A,RPS21 can be used as biomarkers to indicate the malignant event of solely bone marrow relapse,which may be regulated by ZNF460.