Modulation of miR-185-5p expression by EBV-miR-BART6 contributes to developmental differences in ABCG4 gene expression in human megakaryocytes
Modulation of miR-185-5p expression by EBV-miR-BART6 contributes to developmental differences in ABCG4 gene expression in human megakaryocytes
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EBV-miR-BART6对miR-185-5p表达的调节导致人巨核细胞ABCG4基因表达的发育差异
DOI：
10.1016/j.biocel.2016.11.001
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发表时间：
2016-12-01
影响因子：
4
通讯作者：
Hu, Haiyan
中科院分区：
文献类型：
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作者：
Deng, Lan;Wang, Xiuju;Hu, Haiyan
免疫性血小板减少症（ITP）是一种获得性自身免疫性疾病，其特征是血小板计数低和出血，通常由病毒感染引发。我们以前报道过，巨核细胞与ITP患者血清培养的14种病毒microRNA，包括ebv-miR-BART 6，均上调。已有研究报道ebv-miR-BART 6下调miR-185- 5 p的表达。因此，我们预测，ABCG 4基因，这是在巨核细胞祖细胞中高度表达，是miR-185- 5 p的直接目标。我们推测ebv-miR-BART 6可能在巨核细胞的发育和分化中发挥作用。首先，我们通过荧光素酶分析验证了miR-185- 5 p对ABCG 4的负调控。第二，在将ebv-miR-BART 6转染到由正常脐带血单核细胞（MNC）发育而来的巨核细胞中后，我们发现ebv-miR-BART 6组中miR-185- 5 p的水平降低到对照组的近三分之一，伴随着ABCG 4在mRNA和蛋白水平上的上调。同时，与空白对照组和阴性对照组相比，ebv-miR-BART 6组巨核细胞增殖受到明显抑制（分别为14.89% ± 3.13%、34.15% ± 2.42%和30.96% ± 4.37%; P
Immune thrombocytopenia (ITP) is an acquired autoimmune disorder characterized by low platelet count and bleeding, and is usually triggered by viral infections. We previously reported that 14 viral microRNAs of megakaryocytes cultured with serum from patients with ITP, including ebv-miR-BART6, are up-regulated. Previous research has reported that ebv-miR-BART6 down-regulated the expression of miR-185-5p. We therefore predicted that the ABCG4 gene, which is highly expressed in megakaryocyte progenitor cells, is a direct target of miR-185-5p. We hypothesized that ebv-miR-BART6 may play a role in development and differentiation of megakaryocytes. First, we verified the negative regulation of ABCG4 by miR-185-5p through luciferase assay analysis. Second, after transfection of ebv-miR-BART6 into megakaryocytes developing from normal cord blood mononuclear cells (MNCs), we found that the level of miR-185-5p in the ebv-miR-BART6 group was reduced to almost a third of that in the control groups, accompanied by up-regulation of ABCG4 at both the mRNA and protein levels. Meanwhile, proliferation of megakaryocytes was significantly repressed in the ebv-miR-BART6 group compared with the blank and negative control groups (14.89% 3.13%, 34.15% 2.42% and 30.96% 4.37%, respectively; P