Wang Zhong-Dong, Qu Fan-Yong, Chen Yuan-Yuan, Ran Zhang-Shen, Liu Hai-Yan, Zhang Hai-Dong
Clinical Laboratory of Taishan Sanatorium of Shandong Province, Tai'an 271001, China.
Interventional Radiology, Yantai Affiliated Hospital of Binzhou Medical University, Yantai 264100, China.
J Zhejiang Univ Sci B. 2017;18(1):27-36. doi: 10.1631/jzus.B1600205.
Hepatocellular carcinoma (HCC) is still one of the most common death-related malignancies worldwide. Because the way onset and progression are hidden most, HCC diagnoses are made at an advanced stage, when they are unsuitable for surgical resection. MicroRNAs are a class of small non-coding RNAs, participating in many aspects of cancers. In this study, we tried to establish the role of microRNA-718 (miR-718) in the malignant phenotype of HCC cells and its possible role in HCC diagnosis.
Here we first used a methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, Transwell migration and invasion assays, and colony formation assay to evaluate the impact of miR-718 on the malignant phenotypes of HCC cells. Then, we used bioinformatic methods to predict the target gene of miR-718 and used green fluorescence protein (GFP) reporter assay, Western blot, and quantitative real-time polymerase chain reaction (qRT-PCR) to validate the regulation relationship. Finally, we determined the role of the target gene in the HCC phenotype.
We found that the expression of miR-718 was significantly reduced in various HCC cell lines and HCC tissues. Re-expression of miR-718 significantly reduced the cellular viability and colony formation ability as well as inhibited the migration and invasion abilities of HCC cell lines. Early growth response protein 3 (EGR3) is a direct target of miR-718 and is negatively regulated by miR-718. EGR3 could increase the viability and proliferation of HCC cells, and promot the migration and invasion of HCC cells.
miR-718 acts as a tumor suppressive microRNA in HCC via regulating the expression of EGR3, which may provide a new diagnostic marker and treatment target for HCC.
肝细胞癌（HCC）仍是全球最常见的与死亡相关的恶性肿瘤之一。由于其发病和进展方式大多隐匿，HCC往往在不适于手术切除的晚期才得以诊断。微小RNA是一类小的非编码RNA，参与癌症的诸多方面。在本研究中，我们试图确定微小RNA - 718（miR - 718）在HCC细胞恶性表型中的作用及其在HCC诊断中的可能作用。
我们首先使用甲基噻唑基二苯基溴化四氮唑（MTT）法、Transwell迁移和侵袭试验以及集落形成试验来评估miR - 718对HCC细胞恶性表型的影响。然后，我们使用生物信息学方法预测miR - 718的靶基因，并使用绿色荧光蛋白（GFP）报告基因试验、蛋白质免疫印迹法和定量实时聚合酶链反应（qRT - PCR）来验证调控关系。最后，我们确定靶基因在HCC表型中的作用。
我们发现miR - 718在各种HCC细胞系和HCC组织中的表达显著降低。miR - 718的重新表达显著降低了细胞活力和集落形成能力，并抑制了HCC细胞系的迁移和侵袭能力。早期生长反应蛋白3（EGR3）是miR - 718的直接靶标，且受miR - 718负调控。EGR3可增加HCC细胞的活力和增殖，并促进HCC细胞的迁移和侵袭。
miR - 718通过调控EGR3的表达在HCC中发挥肿瘤抑制性微小RNA的作用，这可能为HCC提供一种新的诊断标志物和治疗靶点。
