Comprehensive profiling of novel microRNA-9 targets and a tumor suppressor role of microRNA-9 via targeting IGF2BP1 in hepatocellular carcinoma.
Comprehensive profiling of novel microRNA-9 targets and a tumor suppressor role of microRNA-9 via targeting IGF2BP1 in hepatocellular carcinoma.
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全面分析新型 microRNA-9 靶标以及 microRNA-9 通过靶向 IGF2BP1 在肝细胞癌中的肿瘤抑制作用
DOI：
10.18632/oncotarget.5969
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发表时间：
2015-12-08
期刊：
影响因子：
--
通讯作者：
Lu F
中科院分区：
文献类型：
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作者：
Zhang J;Cheng J;Zeng Z;Wang Y;Li X;Xie Q;Jia J;Yan Y;Guo Z;Gao J;Yao M;Chen X;Lu F
microRNA-9（miR-9）失调与包括肝细胞癌（HCC）在内的多种人类恶性肿瘤有关，但其作用仍然相互矛盾。本研究旨在探讨miR-9在肝癌组织中的表达和甲基化状态，以及miR-9在体外的肿瘤相关功能。通过生物信息学分析、基于芯片的RNA表达谱分析和文献检索来确定HCC中miR-9的靶点。然后通过荧光素酶报告基因测定、实时定量PCR和蛋白质印迹或酶联免疫吸附测定（ELISA）验证潜在的下游候选物。进一步探讨miR-9靶基因在临床标本中的表达状况及临床病理意义。结果显示，miR-9在原发性HCC中频繁下调。它的沉默主要是由高频率（42.5%）的mir-9-1甲基化，这与较大的肿瘤大小（P = 0.0234）。体外功能研究表明，miR-9恢复延缓了HCC细胞的增殖和迁移。IL-6、AP 3B 1、TC 10、ONECUT 2、IGF 2BP 1、MYO 1D和ANXA 2被证实是HCC中的miR-9靶点。其中，ONECUT 2、IGF 2BP 1和ANXA 2在HCC中被证实异常上调。此外，ONECUT 2、IGF 2BP 1和IL-6的上调与术后预后不良显著相关（分别为P = 0.0458、P = 0.0037和P = 0.0461）。在机制上，miR-9部分通过功能性miR-9/IGF 2BP 1/AKT&ERK轴发挥肿瘤抑制作用。我们的研究表明，miR-9通过抑制一系列靶基因，包括新验证的miR-9/IGF 2BP 1/AKT&ERK轴，在HCC进展中发挥肿瘤抑制剂的作用，从而为HCC患者提供潜在的治疗靶点和新的预后生物标志物。
MicroRNA-9 (miR-9) dysregulation is implicated in a variety of human malignancies including hepatocellular carcinoma (HCC), but its role remains contradictory. In this study, we explored the expression and methylation status of miR-9 in HCC samples, as well as the tumor-related functions of miR-9 in vitro. Bioinformatics analysis, array-based RNA expression profile, and literature retrieval were used to identify miR-9 targets in HCC. The potential downstream candidates were then validated by luciferase reporter assay, real-time quantitative PCR, and western blot or enzyme linked immunosorbent assay (ELISA). The expression status and clinicopathologic significances of miR-9 target genes in clinical samples were further explored. The results showed that miR-9 was frequently downregulated in primary HCC. Its silencing was largely contributed by a high frequency (42.5%) of mir-9-1 hypermethylation, which was correlated with bigger tumor size (P = 0.0234). In vitro functional studies revealed that miR-9 restoration retarded HCC cell proliferation and migration. IL-6, AP3B1, TC10, ONECUT2, IGF2BP1, MYO1D, and ANXA2 were confirmed to be miR-9 targets in HCC. Among them, ONECUT2, IGF2BP1, and ANXA2 were confirmed to be aberrantly upregulated in HCC. Moreover, upregulation of ONECUT2, IGF2BP1, and IL-6 were significantly associated with poor post-surgery prognosis (P = 0.0458, P = 0.0037 and P = 0.0461, respectively). Mechanically, miR-9 plays a tumor suppressive role partially through a functional miR-9/IGF2BP1/AKT&ERK axis. Our study suggests that miR-9 functions as a tumor suppressor in HCC progression by inhibiting a series of target genes, including the newly validated miR-9/IGF2BP1/AKT&ERK axis, thus providing potential therapeutic targets and novel prognostic biomarkers for HCC patients.