中国癌症杂志 ›› 2018, Vol. 28 ›› Issue (12): 888-894.doi: 10.19401/j.cnki.1007-3639.2018.12.002

• 论著 • 上一篇    下一篇

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

曹晶珠1,郑 浩2,陶元平2,姚乃心3,黄智平4,汪珍光2,黄 勤1   

  1. 1. 海军军医大学附属长海医院内分泌科,上海 200433 ;
    2. 海军军医大学附属东方肝胆外科医院肝外三科,上海 200433 ;
    3. 海军军医大学基础医学院,上海 200433 ;
    4. 广州军区广州总医院肝胆外科,广东 广州510010
  • 出版日期:2018-12-30 发布日期:2019-01-11
  • 通信作者: 黄 勤 E-mail: qxinyi1220@163.com
  • 基金资助:
    国家自然基金面上项目(81471038);上海市卫生和计划生育委员会科研课题青年项目(20154Y0083)。

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

CAO Jingzhu1, ZHENG Hao2, TAO Yuanping2, YAO Naixin3, HUANG Zhiping4, WANG Zhenguang2,HUANG Qin1   

  1. 1. Endocrine Department of Changhai Hospital, Naval Military Medical University, Shanghai 200433, China; 2. Third Department of Hepatic Surgery, Eastern Hepatobiliary Surgery Hospital, Naval Military Medical University, Shanghai 200433, China; 3. College of Basic Medicine, Naval Military Medical University, Shanghai 200433, China; 4. Department of Hepatobiliary Surgery, General Hospital of Guangzhou Military Command of PLA, Guangzhou 510010, Guangdong
    Province, China
  • Published:2018-12-30 Online:2019-01-11
  • Contact: HUANG Qin E-mail:qxinyi1220@163.com

摘要: 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。

关键词: 肝细胞癌, 微小RNA, 肿瘤转移, 侵袭

Abstract: 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.

Key words: Hepatocellular carcinoma, MicroRNA, Tumor invasion, Metastasis