Lu Caicheng, Liao Zengwei, Cai Minxian, Zhang Guirong
Department of Medical Technology, The Second Hospital of Longyan, Longyan, Fujian 364000, P.R. China.
Oncol Lett. 2017 Feb;13(2):573-578. doi: 10.3892/ol.2016.5479. Epub 2016 Dec 12.
MicroRNAs (miRs) have emerged as key epigenetic regulators involved in cancer progression. miR-320a has been demonstrated to be a novel tumor suppressive microRNA in several types of cancers. In the present study, the role of miR-320a in human hepatocellular carcinoma (HCC) was investigated. The expression levels of miR-320a and messenger RNA were determined by reverse transcription-quantitative polymerase chain reaction, while cell cycle and cell apoptosis were analyzed by flow cytometry. The cell proliferative ability was determined by Cell Counting Kit-8 assay and colony formation assay. The downstream target of miR-320a was confirmed by luciferase reporter assay, while the protein levels were measured by western blotting. The results revealed that miR-320a was inversely associated with HCC proliferation in HCC cell lines. Functional studies demonstrated that miR-320a significantly decreased the capability of cell proliferation and induced G/G growth arrest . In addition, β-catenin was identified as one of the direct targets of miR-320a, downregulating the expression level of β-catenin, c-myc, cyclin D1 and dickkopf-1. In conclusion, miR-320a may act as a tumor-suppressive microRNA through targeting β-catenin in HCC.
微小RNA（miRs）已成为参与癌症进展的关键表观遗传调节因子。miR-320a已被证明是几种癌症中的一种新型肿瘤抑制性微小RNA。在本研究中，对miR-320a在人类肝细胞癌（HCC）中的作用进行了研究。通过逆转录定量聚合酶链反应测定miR-320a和信使RNA的表达水平，同时通过流式细胞术分析细胞周期和细胞凋亡。通过细胞计数试剂盒-8测定法和集落形成测定法确定细胞增殖能力。通过荧光素酶报告基因测定法确认miR-320a的下游靶标，同时通过蛋白质印迹法测量蛋白质水平。结果显示，在肝癌细胞系中，miR-320a与肝癌增殖呈负相关。功能研究表明，miR-320a显著降低细胞增殖能力并诱导G/G生长停滞。此外，β-连环蛋白被确定为miR-320a的直接靶标之一，下调β-连环蛋白、c-myc、细胞周期蛋白D1和Dickkopf-1的表达水平。总之，miR-320a可能通过靶向肝癌中的β-连环蛋白而作为一种肿瘤抑制性微小RNA。
