Gao Bo, Wang Duanping, Ma Wenchao, Jia Xu, Ma Biao, Zhang Weihui, Xue Dongbo
Department of General Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
College of Bioinformatics Science and Technology, Harbin Medical University, Harbin, China.
Clin Res Hepatol Gastroenterol. 2014 Sep;38(4):451-61. doi: 10.1016/j.clinre.2013.09.005. Epub 2014 Jun 24.
As important components of bone marrow microenvironment, bone marrow endothelial cells (BMECs) have important roles in regulating haematopoietic functions of the bone marrow. In preliminary study, we found the humoral inhibitor in liver cirrhosis (LC) could lead to ultrastructure alterations of the bone marrow endothelium. The present study aimed to investigate functional changes occurred in BMECs during LC.
A multi-step approach combining microarray microRNA (miRNA) and mRNA expression profiling and bioinformatics analysis was used to generate a specific miRNA-mRNA regulatory network in BMECs treated with supplemented medium with 20% pooled sera from 26 patients with LC or 10 healthy volunteers as a control group for 48h.
A total of 372 differentially expressed miRNAs and 1872 differentially expressed mRNAs were identified by miRNA and mRNA microarray, respectively. Twenty-one dysregulated miRNAs and their 23 dysregulated target mRNAs confirmed by experiment (31 miRNA-mRNA interactions) were screened based on databases miRecords and miRTarBase. Bioinformatics analysis revealed that the functions of these miRNA-mRNA interactions were involved in the positive regulation of apoptosis, negative regulation of cell proliferation, cell cycle arrest and so on. The results of flow cytometry analysis showed that the LC serum treated-BMECs had a higher apoptosis percentage (17.57±1.84%) compared to the control group (10.2±1.55%). (P<0.05) CONCLUSIONS: Thirty-one miRNA-mRNA pairs combined with 21 dysregulated miRNAs and their 23 target mRNAs identified by the multi-step approach are involved in BMECs treated with LC serum. The results indicated that LC serum-induced apoptosis in BMECs were regulated by a complicated miRNA-mRNA regulatory network.
作为骨髓微环境的重要组成部分，骨髓内皮细胞（BMECs）在调节骨髓造血功能中发挥着重要作用。在前期研究中，我们发现肝硬化（LC）患者体内的体液抑制因子可导致骨髓内皮细胞超微结构改变。本研究旨在探讨肝硬化过程中BMECs发生的功能变化。
采用微阵列微小RNA（miRNA）和信使核糖核酸（mRNA）表达谱分析以及生物信息学分析相结合的多步骤方法，构建用来自26例肝硬化患者的混合血清或10名健康志愿者血清（作为对照组）补充的培养基处理48小时的BMECs中的特定miRNA-mRNA调控网络。
通过miRNA和mRNA微阵列分别鉴定出372个差异表达的miRNA和1872个差异表达的mRNA。基于miRecords和miRTarBase数据库筛选出经实验确认的21个失调miRNA及其23个失调靶mRNA（31个miRNA-mRNA相互作用）。生物信息学分析表明，这些miRNA-mRNA相互作用的功能涉及细胞凋亡的正调控、细胞增殖的负调控、细胞周期阻滞等。流式细胞术分析结果显示，与对照组（10.2±1.55%）相比，LC血清处理的BMECs凋亡率更高（17.57±1.84%）。（P<0.05）结论：通过多步骤方法鉴定出的31对miRNA-mRNA组合以及21个失调miRNA及其23个靶mRNA参与了LC血清处理的BMECs。结果表明，LC血清诱导的BMECs凋亡受复杂的miRNA-mRNA调控网络调节。
