慢性髓系白血病疾病进展相关基因的生物信息学分析与初步实验验证

Bioinformatic Analysis of Chronic Myeloid Leukemia Progression and Preliminary Experimental Verification

本研究探讨慢性髓系白血病(CML)的疾病进展机制,为该疾病临床转归的评估及治疗选择提供新的分子标志物。从NCBI数据库GEO下载一组与CML疾病进展相关的基因芯片数据,利用生物信息学工具分析并得到与加速期(AP)和急变期(BC)相关的基因集,将这两组基因集进行信号通路分析和网络图谱分析得到与进展相关的关键基因和信号通路,并应用荧光定量PCR方法对其中的关键基因进行初步实验验证。结果表明,将AP相关基因和BC相关基因交叉比较,得到在AP和BC都有异常表达的9个基因;信号通路分析发现,整合素介导的细胞黏附,黏着斑细胞粘附以及细胞骨架调节在AP和BC始终发生异常;网络构建分析发现,关键节点的基因分别是MLLT4,WDR35和EPHB4。应用荧光定量PCR方法对疾病进展期和慢性期的患者进行比较发现,EPHB4在进展期的表达水平显著高于慢性期。结论:发现了与CML进展相关的9个基因,其中MLLT4,WDR35,EPHB4基因可能是导致CML进展的关键基因。

This study was aimed to explore the progression mechanism of chronic myeloid leukemia, so as to pro- vide the new molecular markers for evaluation of CML clinical outcome and selection of treatment. The microarray data of genes related with progression from different phase of chronic myeloid leukemia（CML） were collected from public data depository GEO（Gene expression datasets）. SAM analysis, fold change filtering, cross comparison were used to analyze the data and identify different genes. Moreover, MeV and pSTIING sofewares were used to analyze the key dif- ferential genes and signal pathways. At last, Q-PCR were used to confirm the predicted key gene. The results indiated that after comparison, 9 genes were differentially expressed from AP to BC, and the integrin-mediated cell adhesion , fo- cal adhesion, regulation of actin cytoskeleton were the principal pathways during CML progression. Network construction analysis found that AP-related genes or pathways may be the original signals ; and MLLT4, WDR35 and EPHB4 were the key genes for CML progression. EPHB4 was confirmed by Q-PCR in CML BC patients and CP patients. It is concluded that MLLT4, WDR35, EPHB4, integrin-mediated cell adhesion, focal adhesion and regulation of actin cytoskeleton are the principal genes and pathways during CML progression.