Sun Shijie, Wang Ning, Sun Ziwen, Wang Xuefeng, Cui Hongwei
Department of Hepatobiliary Surgery, Yantai Yuhuangding Hospital, Yantai, China.
J BUON. 2019 Jan-Feb;24(1):178-186.
Hepatocellular carcinoma (HCC) is a member of the most frequent malignancies in the world and the poor prognosis of HCC is mainly due to lack of early detection and treatment. The purpose of this study was to investigate the role of microRNA (miR)-5692a in the progression of HCC and its underlying mechanism.
The relative expression of miR-5692a in HCC tissues and cell lines was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) assay. Cell counting kit-8 assay and colony formation assay were used to determine cell proliferation. Flow cytometric analysis was carried out to determine cell cycle distribution and apoptotic cells. Bioinformatics analysis and dual luciferase reporter assay were employed to predict and verify the potential targets of miR-5692a. Protein expression level of HOXD8 was assessed by western blotting normalized by GAPDH in transfected cells.
The relative expression level of miR-5692a was increased in both HCC tissues and cell lines. According to CCK8 assay and colony formation assay, miR-5692a was considered to promote proliferation in HCC. The consequence of flow cytometric analysis showed that overexpressed miR-5692a accelerated cell cycle and inhibited cell apoptosis. We verified that HOXD8 was a target of miR-5692a via online prediction database and dual luciferase reporter assay. The rescue assay we carried out subsequently validated that miR-5692a functioned as an oncogene by regulating HOXD8 in HCC.
This study revealed that miR-5692a had an oncogenic role in HCC by targeting HOXD8 which might bring a novel insight into new therapeutic targets and biomarkers in HCC.
肝细胞癌(HCC)是全球最常见的恶性肿瘤之一,HCC预后较差主要是由于缺乏早期检测和治疗。本研究旨在探讨微小RNA(miR)-5692a在HCC进展中的作用及其潜在机制。
采用定量实时聚合酶链反应(qRT-PCR)检测miR-5692a在HCC组织和细胞系中的相对表达。使用细胞计数试剂盒-8检测法和集落形成检测法来确定细胞增殖情况。通过流式细胞术分析来确定细胞周期分布和凋亡细胞。采用生物信息学分析和双荧光素酶报告基因检测来预测和验证miR-5692a的潜在靶标。通过蛋白质免疫印迹法,以甘油醛-3-磷酸脱氢酶(GAPDH)为内参,评估转染细胞中HOXD8的蛋白表达水平。
miR-5692a在HCC组织和细胞系中的相对表达水平均升高。根据CCK8检测法和集落形成检测法,miR-5692a被认为可促进HCC细胞增殖。流式细胞术分析结果显示,过表达miR-5692a可加速细胞周期进程并抑制细胞凋亡。通过在线预测数据库和双荧光素酶报告基因检测,我们验证了HOXD8是miR-5692a的靶标。随后进行的挽救实验证实,miR-5692a在HCC中通过调控HOXD8发挥癌基因作用。
本研究表明,miR-5692a通过靶向HOXD8在HCC中发挥致癌作用,这可能为HCC的新治疗靶点和生物标志物带来新的见解。