Chen Shu-Ying, Ma De-Ning, Chen Qiu-Dan, Zhang Jing-Jun, Tian Yue-Ru, Wang Zhi-Cheng, Cai Hao, Lin Yong, Sun Hui-Chuan
Huashan Hospital, Fudan University, 12 Wulumuqi middle Road, Shanghai 200040, People's Republic of China.
Department of Liver Surgery, Fudan University, Shanghai Cancer Center, Cancer Hospital, 270 Dongan Road, Shanghai, People's Republic of China.
J Cancer. 2017 Feb 25;8(4):617-625. doi: 10.7150/jca.17394. eCollection 2017.
MicroRNAs (miRNAs) are a class of endogenous, small non-coding RNAs which function as essential posttranscriptional modulators of gene expression tightly involved in a wide range of diseases, including the hepatocellular carcinoma (HCC). Here, the present study was designed to investigate the expression levels and cellular roles of miR-200a in HCC. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-200a in serums and cell lines. Bioinformation analysis, the luciferase reporter assay, qRT-PCR and western blotting were employed to validate Foxa2 as a direct target gene of miR-200a. Cell proliferation, migration and invasion were assessed to identify whether miR-200a could regulate the biological behaviors of HCC cells by targeting Foxa2. In this study, a low level of miR-200a was observed in patients' serums and HCC cell lines. Overexpression of miR-200a in HCC cell lines reduced cell proliferation, migration and invasion. In addition, transcription factor forkhead box A2 (Foxa2) was identified as a novel target of miR-200a and downregulated at mRNA and protein levels in miR-200a overexpressed cells. Meanwhile, restoration of Foxa2 significantly reversed the tumor suppressive effects of miR-200a. These findings indicate that miR-200a regulates the proliferation, migration and invasion of HCC cells by targeting Foxa2, suggesting that miR-200a may function as a potential therapeutic molecular for the diagnosis and treatment of the liver cancer.
微小RNA（miRNA）是一类内源性的小非编码RNA，作为基因表达的重要转录后调节因子，紧密参与包括肝细胞癌（HCC）在内的多种疾病。在此，本研究旨在探讨miR-200a在肝癌中的表达水平及细胞作用。采用定量逆转录聚合酶链反应（qRT-PCR）检测血清和细胞系中miR-200a的表达水平。运用生物信息分析、荧光素酶报告基因检测、qRT-PCR和蛋白质免疫印迹法验证叉头框蛋白A2（Foxa2）是miR-200a的直接靶基因。通过评估细胞增殖、迁移和侵袭，以确定miR-200a是否可通过靶向Foxa2调节肝癌细胞的生物学行为。在本研究中，观察到患者血清和肝癌细胞系中miR-200a水平较低。在肝癌细胞系中过表达miR-200a可降低细胞增殖、迁移和侵袭能力。此外，转录因子叉头框A2（Foxa2）被鉴定为miR-200a的新靶标，在miR-200a过表达的细胞中其mRNA和蛋白质水平下调。同时，恢复Foxa2表达可显著逆转miR-200a的抑癌作用。这些发现表明，miR-200a通过靶向Foxa2调节肝癌细胞的增殖、迁移和侵袭，提示miR-200a可能作为一种潜在的治疗分子用于肝癌的诊断和治疗。
