UNC5B-AS1 promotes the proliferation, migration and EMT of hepatocellular carcinoma cells via regulating miR-4306/KDM2A axis

Being one of the most prevalent malignancies, hepatocellular carcinoma (HCC) threatens the health of population all over the world. Numerous researches have confirmed that long noncoding RNAs (lncRNAs) play an important role in tumor progression. Nonetheless, the mechanisms of unc-5 netrin receptor B antisense RNA 1 (UNC5B-AS1) in HCC remain obscure. Thus, this study aims to investigate the regulatory role and mechanism of UNC5B-AS1 in HCC cells. In our research, UNC5B-AS1 was subjected to gene expression analysis by RT-qPCR. Biological functions of UNC5B-AS1 in HCC cells were measured by MTT, colony formation, EdU and transwell assays. The combination between UNC5B-AS1, lysine demethylase 2A (KDM2A) and miR-4306 was validated by mechanism assays. Result showed UNC5B-AS1 was upregulated in HCC tissues and cells, contributing to the development of cancer staging and survival rate of HCC patients. Moreover, UNC5B-AS1 deficiency inhibited the proliferation, migration and epithelial–mesenchymal transition (EMT) of HCC cells. Furthermore, UNC5B-AS1 could interact with miR-4306 in HCC cells. Similarly, KDM2A was proved as the target gene of miR-4306. Finally, miR-4306 downregulation or KDM2A overexpression reversed the prohibitive role of UNC5B-AS1 knockdown in HCC progression. In short, UNC5B-AS1 accelerates the proliferation, migration and EMT of HCC cells via the regulation of miR-4306/KDM2A axis.

作为全球范围内最常见的恶性肿瘤之一，肝细胞癌（hepatocellular carcinoma, HCC）严重威胁全球人群的健康。诸多研究已证实，长链非编码RNA（long noncoding RNAs, lncRNAs）在肿瘤进展过程中发挥关键调控作用。然而，unc-5网蛋白受体B反义RNA 1（UNC5B-AS1）在肝细胞癌中的作用机制仍有待阐明。因此，本研究旨在探讨UNC5B-AS1在肝细胞癌细胞中的调控作用及具体分子机制。本研究采用实时荧光定量聚合酶链反应（RT-qPCR）对UNC5B-AS1的基因表达水平进行检测分析。通过MTT实验、平板克隆形成实验、5-乙炔基-2'-脱氧尿苷（EdU）染色实验及Transwell实验，检测UNC5B-AS1在肝细胞癌细胞中的生物学功能。通过机制实验验证了UNC5B-AS1、赖氨酸去甲基化酶2A（lysine demethylase 2A, KDM2A）与miR-4306三者之间的相互结合关系。实验结果显示，UNC5B-AS1在肝细胞癌组织及细胞中呈高表达状态，其表达水平与肝细胞癌患者的临床分期进展及不良生存率密切相关。此外，敲低UNC5B-AS1可显著抑制肝细胞癌细胞的增殖、迁移及上皮间质转化（epithelial–mesenchymal transition, EMT）进程。进一步研究发现，UNC5B-AS1可在肝细胞癌细胞中与miR-4306发生特异性相互作用。同样地，赖氨酸去甲基化酶2A被证实为miR-4306的靶基因。最终实验结果表明，miR-4306表达下调或赖氨酸去甲基化酶2A过表达，可逆转敲低UNC5B-AS1对肝细胞癌进展的抑制作用。综上，UNC5B-AS1可通过调控miR-4306/KDM2A信号轴，加速肝细胞癌细胞的增殖、迁移及上皮间质转化进程。