Sun Jian-Jun, Chen Guo-Yong, Xie Zhan-Tao
Department of Hepatobiliary Surgery, Henan Provincial People's Hospital, Henan, China.
Cell Physiol Biochem. 2016;38(2):777-85. doi: 10.1159/000443033. Epub 2016 Feb 15.
BACKGROUND/AIMS: A growing body of evidence supports the notion that MicroRNAs (miRNAs) function as key regulators of tumorigenesis. In the present study, the expression and roles of miRNA-361-5p were explored in hepatocellular carcinoma (HCC).
Quantitative real-time PCR was used to detect the expression miR-361-5p in HCC tissues and pair-matched adjacent normal tissues. MTT and BrdU assays were used to identify the role of miR-361-5p in the regulation of proliferation and invasion of HCC cells. Using bioinformatics analysis, luciferase reporter assays and Western blots were used to identify the molecular target of miR-361-5p. nude mice were used to detect the anti-tumor role of miR-361-5p in vivo.
miR-361-5p was down-regulated in HCC tissues in comparison to adjacent normal tissues, due to hypermethylation at its promoter region. Overexpression of miR-361-5p suppressed proliferation and invasion of HCC cells. Chemokine (C-X-C Motif) receptor 6 (CXCR6) was identified as a target of miR-361-5p. Indeed, knockdown of CXCR6 photocopied, while overexpression of CXCR6 largely attenuated the anti-proliferative effect of miR-361-5p. More importantly, in vivo studies demonstrated that forced expression of miR-361-5p significantly inhibited tumor growth in the nude mice.
Our results indicate that miR-361-5p acts as a tumor suppressor and might serve as a novel therapeutic target for the treatment of HCC patients.
背景/目的：越来越多的证据支持微小RNA（miRNA）作为肿瘤发生关键调节因子的观点。在本研究中，探讨了miRNA-361-5p在肝细胞癌（HCC）中的表达及作用。
采用定量实时PCR检测HCC组织及配对的癌旁正常组织中miR-361-5p的表达。采用MTT和BrdU检测法确定miR-361-5p在调节HCC细胞增殖和侵袭中的作用。通过生物信息学分析、荧光素酶报告基因检测和蛋白质免疫印迹法确定miR-361-5p的分子靶点。使用裸鼠检测miR-361-5p在体内的抗肿瘤作用。
与癌旁正常组织相比，HCC组织中miR-361-5p表达下调，这是由于其启动子区域的高甲基化所致。miR-361-5p的过表达抑制了HCC细胞增殖和侵袭。趋化因子（C-X-C基序）受体6（CXCR6）被确定为miR-361-5p的靶点。实际上，CXCR6的敲低模拟了miR-361-5p的作用，而CXCR6的过表达则大大减弱了miR-361-5p的抗增殖作用。更重要的是，体内研究表明，miR-361-5p的强制表达显著抑制了裸鼠肿瘤生长。
我们的结果表明，miR-361-5p作为一种肿瘤抑制因子，可能成为治疗HCC患者的新治疗靶点。
