miR-509-3p在肝癌组织中的表达及对肝癌细胞迁移侵袭的影响

背景与目的：肝细胞癌（hepatocellular carcinoma，HCC）是世界范围内致死率第三高的恶性肿瘤，microRNA（miRNA）被认为在HCC的发病机制中起重要作用。本研究旨在探讨在原发性肝癌中miR-509-3p的表达水平、细胞功能及调控的相关基因。方法：通过实时荧光定量聚合酶链反应（real-time fluorescent quantitative polymerase chain reaction，RTFQ-PCR）技术检测miR-509-3p在46例肝癌患者组织样本、人肝癌细胞系及永生化人正常肝细胞系中的表达水平，采用transwell实验检测miR-509-3p对肝癌细胞转移的影响。采用蛋白质印迹法（Western blot）检测miR-509-3p与上皮-间质转化（epithelial-mesenchymal transition，EMT）相关基质金属蛋白酶（matrix metalloproteinase，MMP）之间的关系。结果：RTFQ-PCR技术检测结果显示，miR-509-3p在46例肝癌患者的肝癌组织样本以及MHCC97H、HCCLM3和SMMC-7721肝癌细胞系中异常高表达。体外功能实验证实抑制miR-509-3p表达能降低MHCC97H和HCCLM3肝癌细胞的迁移能力。Western blot检测结果显示，在MHCC97H和HCCLM3肝癌细胞中抑制miR-509-3p的表达可以降低MMP-9和MMP-2蛋白的表达。结论：本研究表明miR-509-3p通过正向调控MMP-9和MMP-2蛋白的表达而促进肝癌细胞的迁移和侵袭。

Expression of miR-509-3p and its effect on migration and invasion of hepatocellular carcinoma

Background and purpose: This study aimed to investigate the expression, cellular functions and mechanisms of miR-509-3p in hepatocellular carcinoma (HCC). Methods: miR-509-3p was detected by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) in an independent validation sample cohort of 46 HCC tissues, 3 HCC cell lines and one normal liver cell line. Transwell assay was performed to evaluate the effect of miR-509-3p on metastasis in HCC cells. Western blot was performed to evaluate the relationship between miR-509-3p and matrix metalloproteinases (MMPs) associated with epithelial-mesenchymal transition (EMT). Results: miR-509-3p was highly expressed in HCC tissues and HCC cell lines. In vitro functional experiments showed that the inhibition of mir-509-3p decreased the metastatic ability of HCC cells. Western blot analysis confirmed that mir-509-3p negatively regulated the protein level of MMP-9 and MMP-2. Conclusion: This study shows that mir-509-3p promotes EMT in HCC cells through specific signaling pathway and molecular mechanisms.