目的 探讨肝细胞癌中miR-3682-3p的表达情况、临床意义和作用机制。方法 待实验所需肝癌细胞、正常肝细胞（L02）培养与转染后，采用实时定量PCR（qPCR）检测miR-3682-3p在肝癌组织、癌旁组织、L02细胞和肝癌细胞系中的表达情况，利用TCGA数据库分析正常肝组织、肝癌组织中miR-3682-3p的表达差异，分析miR-3682-3p的表达与肝癌患者临床病理特征、预后的关系；通过Transwell实验评价细胞侵袭、迁移能力；通过TargetScan数据库预测获得下游靶基因，采用qPCR、Western blotting法分析miR-3682-3p对生长抑制特异性基因8（GAS8）的表达的调控作用，并利用双荧光素酶报告基因实验加以验证。结果 与癌旁组织和正常肝细胞比较，miR-3682-3p在肝癌组织、细胞系中均高表达（P<0.05），并与门静脉侵犯、Edmondson分级、TNM分期有关；基于TCGA数据库数据的生存分析显示，miR-3682-3p高表达与不良预后相关；在MHCC97-H细胞中敲减miR-3682-3p显著抑制细胞的侵袭、迁移（P<0.05）；通过生物信息学预测加以实验验证证实GAS8是miR-3682-3p的直接靶基因，并通过回复实验证实GAS8介导了miR-3682-3p对肝癌细胞侵袭及迁移能力的促进作用。结论 miR-3682-3p在肝癌中通过靶向抑制抑癌基因GAS8从而促进肝癌细胞侵袭、迁移。

Objective To investigate the expression level, clinical significance and mechanisms of miR-3682-3p in human hepatocellular carcinoma (HCC). Methods After the HCC cells and normal hepatocytes (L02) were cultured and transfected, Real-time quantitative PCR (qPCR) was used to detect the expression of miR-3682-3p in HCC tissues and paracancerous tissues, L02normal liver cells, and HCC cell lines. TCGA database was used to analyze miR-3682 in normal liver tissues and liver cancer tissues, and expression difference, analysis of the relationship between miR-3682-3p expression and clinicopathological features and prognosis of patients with HCC. Transwell assay was used to evaluate the invasion and migration ability of cells. The target gene was predicted by TargetScan database, and the regulation of GAR8 expression by miR-3682-3p was analyzed by qPCR and Western blotting, and the dual luciferase reporter gene assay was used. Results Compared with tumor adjacent tissues or normal hepatic cell, miR-3682-3p was increased in HCC tissue and cell lines (P<0.05). The miR-3682-3p expression was significantly related with the portal vein infiltration, advanced Edmondson degree, and advanced TNM stage. The result of overall survival investigated by TCGA database showed the survival rate in patients with high miR-3682-3p expression was significantly lower than that in those with low miR-3682-3p expression (P<0.05). Knockdown of miR-3682-3p significantly inhibited MHCC97-H cell migration and invasion compared with negative controls (P<0.05). GAS8, as the potential downstream of miR-3682-3p, was validated by luciferase reporter assay. Rescue assay illuminated that GAS8 mediates the effects of miR-3682-3p on migration and invasion of HCC cells. Conclusion miR-3682-3p is upregulated in HCC, which promotes HCC cell migration and invasion through by targeting GAS8.

陕西省自然科学基础研究计划项目（2017JM8002）