MicroRNA-145 inhibits the growth, invasion, metastasis and angiogenesis of neuroblastoma cells through targeting hypoxia-inducible factor 2 alpha
MicroRNA-145 inhibits the growth, invasion, metastasis and angiogenesis of neuroblastoma cells through targeting hypoxia-inducible factor 2 alpha
复制标题
MicroRNA-145通过靶向缺氧诱导因子2α抑制神经母细胞瘤细胞的生长、侵袭、转移和血管生成
DOI：
10.1038/onc.2012.574
复制
发表时间：
2014-01-16
期刊：
影响因子：
8
通讯作者：
Tong, Q.
中科院分区：
文献类型：
--
作者：
Zhang, H.;Pu, J.;Tong, Q.
最近的证据表明，缺氧诱导因子2 α (HIF-2 α)可能在非缺氧条件下神经母细胞瘤（NB）的生长和进展中起关键作用。然而，在NB中正常HIF-2 α表达的潜在机制和临床潜力在很大程度上仍然未知。本研究中，在26/42的NB组织中发现HIF-2 α免疫染色，这与临床病理特征相关。在近20例NB病例中，microRNA-145 （miR-145）下调，并与HIF-2 α表达呈负相关。生物信息学分析显示，在HIF-2 α信使RNA （mRNA）的3'-非翻译区（3'-UTR）可能存在miR-145结合位点。在正常培养的NB细胞系SH-SY5Y和SK-N-SH中，miR-145的过表达或敲低反应性地改变了HIF-2 α及其下游基因cyclin D1、基质金属蛋白酶14和血管内皮生长因子的mRNA和蛋白水平。在一个荧光素酶报告系统中，miR-145下调HIF-2 α 3′-UTR的荧光素酶活性，而这些作用被miR-145结合位点的突变所消除。在体外和体内，过表达miR-145可抑制SH-SY5Y和SK-N-SH细胞的生长、侵袭、转移和血管生成，而恢复HIF-2 α表达可使肿瘤细胞摆脱miR-145介导的这些生物学特征缺陷。此外，anti-miR-145抑制剂挽救了HIF-2 α敲低介导的对NB细胞生长、迁移、侵袭和血管生成的抑制。这些数据表明，在常氧条件下，miR-145通过3'-UTR结合位点抑制HIF-2 α的表达，从而抑制NB的侵袭性和血管生成。
Recent evidence shows that hypoxia-inducible factor 2 alpha (HIF-2 alpha) may have critical roles in the growth and progression of neuroblastoma (NB) under non-hypoxic conditions. However, the underlying mechanisms and clinical potentials of normoxic HIF-2 alpha expression in NB still remain largely unknown. In this study, HIF-2 alpha immunostaining was identified in 26/42 NB tissues, which was correlated with clinicopathological features. In subtotal 20 NB cases, microRNA-145 (miR-145) was downregulated and inversely correlated with HIF-2 alpha expression. Bioinformatics analysis revealed a putative miR-145 binding site in the 3'-untranslated region (3'-UTR) of HIF-2 alpha messenger RNA (mRNA). Overexpression or knockdown of miR-145 responsively altered both the mRNA and protein levels of HIF-2 alpha and its downstream genes, cyclin D1, matrix metalloproteinase 14 and vascular endothelial growth factor, in normoxically cultured NB cell lines SH-SY5Y and SK-N-SH. In a luciferase reporter system, miR-145 downregulated the luciferase activity of HIF-2 alpha 3'-UTR, and these effects were abolished by a mutation in the putative miR-145-binding site. Overexpression of miR-145 suppressed the growth, invasion, metastasis and angiogenesis of SH-SY5Y and SK-N-SH cells in vitro and in vivo, while restoration of HIF-2 alpha expression rescued the tumor cells from miR-145-mediated defects in these biological features. Furthermore, anti-miR-145 inhibitor rescued the HIF-2 alpha knockdown-mediated repression on the growth, migration, invasion and angiogenesis of NB cells. These data indicate that miR-145 suppresses HIF-2 alpha expression via the binding site in the 3'-UTR under normoxic conditions, thus inhibiting the aggressiveness and angiogenesis of NB.