Whole genome expression profiling and screening for differentially expressed cytokine genes in human bone marrow endothelial cells treated with humoral inhibitors in liver cirrhosis
Whole genome expression profiling and screening for differentially expressed cytokine genes in human bone marrow endothelial cells treated with humoral inhibitors in liver cirrhosis
复制标题
肝硬化体液抑制剂治疗的人骨髓内皮细胞的全基因组表达谱和差异表达细胞因子基因的筛选。
DOI：
10.3892/ijmm.2013.1495
复制
发表时间：
2013-11-01
影响因子：
5.4
通讯作者：
Xue, Dongbo
中科院分区：
文献类型：
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作者：
Gao, Bo;Sun, Wang;Xue, Dongbo
骨髓内皮细胞（BMEC）是骨髓造血微环境的重要组成部分，能分泌多种细胞因子调节造血干/祖细胞的功能。迄今为止，还不清楚在肝硬化（LC）的情况下，BMEC是否发生功能变化并导致造血异常。在本研究中，全基因组芯片分析进行检测差异表达的基因在人BMEC处理48小时的培养基中补充了20%的混合血清从26例LC或10名健康志愿者作为对照组。共鉴定了1,106个上调基因和766个下调基因。在基因本体分析中，揭示了最重要的基因类别。大量上调的基因参与细胞间粘附、细胞凋亡和细胞对刺激的反应等过程，而下调的基因则参与分泌、血管生成、血管发育和细胞生长等过程的负调控。通路分析显示，上调表达的基因为细胞粘附分子或凋亡信号通路的一部分，下调表达的基因涉及Wnt信号通路和MAPK信号通路。这些是富集分数最高的途径。细胞凋亡检测结果显示，LC患者血清中的体液抑制因子可诱导BMEC凋亡，证实了生物信息学分析的准确性。筛选并验证了21个差异表达的细胞因子基因[转化生长因子（TGF）B1、肿瘤坏死因子（TNF）B、TNF受体超家族成员11 B（TNFRSF 11B），TNF（配体）超家族，成员13 b（TNFSF 13 B），白细胞介素（IL）1A，IL 6，IL 11，IL 17 C，IL 24，具有序列相似性的家族3，成员B（FAM 3 B），Fas配体（FASLG），基质金属肽酶（MMP）3，MMP 15，玻连蛋白（VTN）、胰岛素样生长因子1（IGF 1）、成纤维细胞生长因子22（FGF 22）、狭缝同源物2（果蝇）（SLIT 2）、血小板反应蛋白（THBS）2、THBS 3、趋化因子（C-C基序）配体28（CCL 28）和巨噬细胞刺激因子1（MST 1）]。我们的研究结果表明，LC患者血清中的体液抑制剂诱导BMEC功能障碍和细胞因子分泌异常，这可能是导致LC患者血液学异常的一种新机制。
Bone marrow endothelial cells (BMECs) are important components of the hematopoietic microenvironment in bone marrow, and they can secrete several types of cytokines to regulate the functions of hematopoietic stem/progenitor cells. To date, it is unknown whether BMECs undergo functional changes and lead to hematopoietic abnormalities in cases of liver cirrhosis (LC). In the present study, whole genome microarray analysis was carried out to detect differentially expressed genes in human BMECs treated for 48 h with medium supplemented with 20% pooled sera from 26 patients with LC or 10 healthy volunteers as the control group. A total of 1,106 upregulated genes and 766 downregulated genes were identified. In Gene Ontology analysis, the most significant categories of genes were revealed. A large number of the upregulated genes were involved in processes, such as cell-cell adhesion, apoptosis and cellular response to stimuli and the downregulated genes were involved in the negative regulation of secretion, angiogenesis, blood vessel development and cell growth. Pathway analysis revealed that the upregulated genes were either cell adhesion molecules or parts of the apoptotic signaling pathway and the downregulated genes were involved in the Wnt signaling pathway and MAPK signaling pathway. These were the pathways with the highest enrichment scores. The results of apoptosis assays revealed that the humoral inhibitors in the sera of patients with LC induced the apoptosis of BMECs, which confirmed the accuracy of bioinformatic analysis. Moreover, we screened and verified 21 differentially expressed cytokine genes [transforming growth factor (TGF)B1, tumor necrosis factor (TNF)B, TNF receptor superfamily, member 11b (TNFRSF11B), TNF (ligand) superfamily, member 13b (TNFSF13B), interleukin (IL)1A, IL6, IL11, IL17C, IL24, family with sequence similarity 3, member B (FAM3B), Fas ligand (FASLG), matrix metallopeptidase (MMP)3, MMP15, vitronectin (VTN), insulin-like growth factor 1 (IGF1), fibroblast growth factor 22 (FGF22), slit homolog 2 (Drosophila) (SLIT2), thrombospondin (THBS)2, THBS3, chemokine (C-C motif) ligand 28 (CCL28) and macrophage stimulating 1 (MST1)] from 97 cytokine genes in BMECs treated with serum from patients with LC. The results from our study demonstrate that the humoral inhibitors in the sera of patients with LC induce the dysfunction and abnormal cytokine secretion by BMECs, which may be a novel mechanism responsible for hematological abnormalities in patients with LC.