microRNA-23b suppresses epithelial-mesenchymal transition (EMT) and metastasis in hepatocellular carcinoma via targeting Pyk2
microRNA-23b suppresses epithelial-mesenchymal transition (EMT) and metastasis in hepatocellular carcinoma via targeting Pyk2
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DOI：
10.1016/j.biopha.2017.02.030
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发表时间：
2017-05-01
影响因子：
7.5
通讯作者：
Yan, Yiqun
中科院分区：
文献类型：
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作者：
Cao, Jie;Liu, Jinkai;Yan, Yiqun
许多microRNA（miRNAs）已被证明在各种癌症中发挥重要作用，包括肝细胞癌（HCC）。然而，miRNAs在肝癌发生、发展和转移中的作用机制仍不清楚。我们从GEO数据库（GSE 26323）中下载了原发性肝癌和肺转移癌的microRNA表达谱的标准化数据，获得了一组转移相关的候选miRNAs。在这些候选miRNAs中，我们重点对miR-23 b进行了进一步的研究。还探索了转移相关的miR-23 b与生存率的关系。通过MTT增殖实验、伤口愈合实验和迁移实验检测miR-23 b在HCC中的生物学作用，并通过Western blot检测EMT相关标志物。利用TargetScan和PicTar预测miR-23 b的潜在靶基因，并通过荧光素酶活性测定进行验证。进行拯救实验以验证miR-23 b的功能是否通过调节其靶标来发挥。我们的研究结果表明，miR-23 b在HCC组织中的表达显著降低，更重要的是，与HCC的肝内转移呈正相关。同时，miR-23 b低表达患者的预后明显较差。过表达miR-23 b可抑制MHCC 97 L细胞的增殖、迁移、侵袭，并调节MMPs和EMT相关基因的表达。此外，Pyk 2是EMT的关键调节因子之一，被鉴定为miR-23 b的直接靶点。此外，Pyk 2过表达可恢复miR-23 b过表达对肿瘤转移的抑制作用。这项研究表明，miR-23 b是一种肿瘤抑制因子，可能通过调节EMT靶向Pyk 2来调节HCC的迁移和侵袭，暗示了HCC治疗的潜在预后生物标志物和治疗靶点。(C)2017 Elsevier Masson SAS。All rights reserved.
Numerous microRNAs (miRNAs) have been shown to play important roles in various cancers, including hepatocellular carcinoma (HCC). However, the functions and mechanisms of the miRNAs involved in HCC progress and metastasis still remain unknown. We downloaded the normalized data of microRNA expression profiling of HCC comparing primary tumor with lung metastasis from GEO database (GSE26323), and gain a group of metastasis-related candidate miRNAs. Among the candidate miRNAs, we focused on miR-23b for further study. The association of metastasis-related miR-23b with survival was also explored. Furthermore, the effects of miR-23b on biological role in HCC were demonstrated by MTT proliferation assay, wound healing and migration assay and the EMT related markers was analyzed by Western blot. Potential target genes of miR-23b were predicted using TargetScan and PicTar and confirmed by luciferase activity assay. A rescue experiment was performed to verify whether the function of miR-23b was exerted via regulation of its target. Our results showed that miR-23b expression was significantly decreased in HCC tissues, which was more importantly, positively correlated to the intrahepatic metastasis of HCC. Meanwhile, patients with low miR-23b expression had significantly poorer prognosis. Overexpression of miR-23b could inhibit MHCC97L cell proliferation, migration, invasion and regulate the expression of MMPs and EMT-associated genes. Moreover, Pyk2, one of the crucial regulators of EMT, was identified as a direct target of miR-23b. In addition, the inhibitory effects of miR-23b overexpression on the metastasis could be restored by Pyk2 overexpression. This study revealed that miR-23b was a tumor suppressor which may regulate HCC migration and invasion by targeting Pyk2 via regulation of EMT, implicating a potential prognostic biomarker and therapeutic target for HCC treatment. (C) 2017 Elsevier Masson SAS. All rights reserved.