Long noncoding RNA H19 upregulates vascular endothelial growth factor A to enhance mesenchymal stem cells survival and angiogenic capacity by inhibiting miR-199a-5p
Long noncoding RNA H19 upregulates vascular endothelial growth factor A to enhance mesenchymal stem cells survival and angiogenic capacity by inhibiting miR-199a-5p
复制标题
长非编码RNA H19通过抑制miR-199a-5p上调血管内皮生长因子A以增强间充质干细胞的存活和血管生成能力
DOI：
10.1186/s13287-018-0861-x
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发表时间：
2018-04-19
影响因子：
7.5
通讯作者：
Wang, Tong
中科院分区：
文献类型：
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作者：
Hou, Jingying;Wang, Lingyun;Wang, Tong
目前，间充质干细胞（MSCs）移植治疗心血管疾病的整体疗效并不令人满意。移植细胞在局部梗死组织中的低活力和血管生成能力限制了它们的进一步应用。有证据表明，长链非编码RNA H19 （lncRNA-H19）介导细胞存活和血管生成。此外，它还参与MSCs的生物活性。本研究旨在探讨lncRNA-H19在MSCs存活和血管生成能力中的功能作用及其机制。方法从C57BL/6小鼠身上获得smscs并进行体外培养。第三代细胞分为MSCs+H19、MSCs+H19 NC、MSCs+si-H19、MSCs+si-H19 NC和MSCs。MSCs+H19和MSCs+H19 NC组分别转染lncRNA-H19和lncRNA-H19 scramble RNA。MSCs+si-H19和MSCs+si-H19 NC组分别转染lncRNA-H19 siRNA和lncRNA-H19 siRNA scramble。以MSCs作为空白对照。所有组均暴露于常氧（20% O2）和缺氧(1% O2)/血清剥夺（H/SD）条件下24 H。评估细胞增殖、凋亡和血管密度。进行生物信息学和双荧光素酶报告基因检测。在不同的实验组中检测相关的生物标志物。结果lncRNA-H19过表达可提高MSCs在常氧和H/SD条件下的存活率和血管生成能力，而lncRNA-H19过表达可降低MSCs的生存能力和血管生成潜能。MicroRNA-199a-5p （miR-199a-5p）靶向并下调血管内皮生长因子A （VEGFA）。MiR-199a-5p是lncRNA-H19的靶标。LncRNA-H19转染导致miR-199a-5p水平降低，同时VEGFA表达升高。然而，在lncRNA-H19敲低的情况下，miR-199a-5p和VEGFA均呈现相反的变化。结论slncrna - h19增强了MSCs的体外存活和血管生成潜能。它可以通过抑制miR-199a-5p作为竞争内源性RNA直接上调VEGFA的表达。这一机制有助于更好地了解MSCs的生物学活性，并为基于MSCs移植的细胞治疗提供新的见解。
BackgroundCurrently, the overall therapeutic efficiency of mesenchymal stem cells (MSCs) transplantation for the treatment of cardiovascular disease is not satisfactory. The low viability and angiogenic capacity of the implanted cells in the local infarct tissues restrict their further application. Evidence shows that long noncoding RNA H19 (lncRNA-H19) mediates cell survival and angiogenesis. Additionally, it is also involved in MSCs biological activities. This study aimed to explore the functional role of lncRNA-H19 in MSCs survival and angiogenic capacity as well as the underlying mechanism.MethodsMSCs were obtained from C57BL/6 mice and cultured in vitro. Cells at the third passage were divided into the following groups: MSCs+H19, MSCs+H19 NC, MSCs+si-H19, MSCs+si-H19 NC and MSCs. The MSCs+H19 and MSCs+H19 NC groups were transfected with lncRNA-H19 and lncRNA-H19 scramble RNA respectively. The MSCs+si-H19 and MSCs+si-H19 NC groups were transfected with lncRNA-H19 siRNA and lncRNA-H19 siRNA scramble respectively. MSCs were used as the blank control. All groups were exposed to normoxia (20% O2) and hypoxia (1% O2)/serum deprivation (H/SD) conditions for 24 h. Cell proliferation, apoptosis and vascular densities were assessed. Bioinformatics and dual luciferase reporter assay were performed. Relevant biomarkers were detected in different experimental groups.ResultsOverexpression of lncRNA-H19 improved survival and angiogenic capacity of MSCs under both normoxia and H/SD conditions, whereas its knockdown impaired cell viability and their angiogenic potential. MicroRNA-199a-5p (miR-199a-5p) targeted and downregulated vascular endothelial growth factor A (VEGFA). MiR-199a-5p was a target of lncRNA-H19. LncRNA-H19 transfection led to a decreased level of miR-199a-5p, accompanied with an elevated expression of VEGFA. However, both miR-199a-5p and VEGFA presented inverse alterations in the condition of lncRNA-H19 knockdown.ConclusionsLncRNA-H19 enhanced MSCs survival and their angiogenic potential in vitro. It could directly upregulate VEGFA expression by inhibiting miR-199a-5p as a competing endogenous RNA. This mechanism contributes to a better understanding of MSCs biological activities and provides new insights for cell therapy based on MSCs transplantation.