目的：探讨miR-377-5p 与缺氧诱导因子-1α（hypoxia inducible factor-1，HIF-1α）的靶向关系以及通过控血管内皮生长因子（vascular endothelial growth factor, VEGF）信号通路对肝细胞癌（hepatocellular carcinoma，HCC）细胞增殖、侵袭和EMT的调控作用。方法：qPCR检测35 例人HCC组织及癌旁组织标本中miR-377-5p 的表达水平。将HepG2 细胞分为对照组、mimic NC组、miR-377-5p mimic 组，qPCR 检测转染效率；EdU染色、Transwell 和Western blotting（WB）检测miR-377-5p 过表达对HepG2 细胞增殖、侵袭及其增殖相关蛋白Ki-67、增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）及上皮间质转化（epithelial-mesenchymaltransition，EMT）标志蛋白E-cadherin、N-cadherin 表达的影响；qPCR、WB检测miR-377-5p 过表达对HepG2 细胞中，缺氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）表达的影响。荧光素酶报告基因实验验证miR-377-5p 与HIF-1α 基因的靶向关系。结果：HCC 组织中miR-377-5p 表达水平较癌旁组织降低（P<0.01）。与对照组相比，miR-377-5p mimic 组HepG2细胞中miR-377-5p 水平明显升高，细胞的增殖、侵袭能力降低（均P<0.01），EMT、N-cadherin 表达水平降低（均P<0.01）而E-cadherin 表达水平显著升高（P<0.01）；miR-377-5p mimic 组中HIF-1α mRNA 和蛋白表达水平均降低（P<0.01 或P<0.05）。miR-377-5p 靶向抑制HIF-1α 基因表达，并抑制VEGF通路的激活（均P<0.05）。结论：miR-377-5p 通过靶向抑制HIF-1α 基因表达和下调VEGF信号通路从而抑制HepG2 细胞的增殖、侵袭和EMT。

Objective：To explore the targeting relationship between miR-377-5p and hypoxia inducible factor-1 (HIF-1α), and investigate the regulatory effect of miR-377-5p on proliferation, invasion and epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma (HCC) cells through vascular endothelial growth factor (VEGF) signaling pathway. Methods：The expression of miR-377-5p in 35 pairs of human HCC tissues and para-cancerous tissues was detected by qPCR. Then, HepG2 cells were divided into control group, mimic-NC group and miR-377-5p mimic group. qPCR was used to detect the transfection efficiency; the effects of miR-377-5p over-expression on proliferation and invasion of HepG2 cells were examined by EdU staining and Transwell assay, respectively; and the effect of miR-377-5p over-expression on the expressions of proliferation-related protein Ki-67, proliferating cell nuclear antigen (PCNA) and epithelial-mesenchymal transition (EMT) markers (E-cadherin and N-cadherin) were detected by Western blotting (WB);the effect of miR-377-5p over-expression on the expression of hypoxia inducible factor-1α (HIF-1α ) in HepG2 cells was detected by qPCR and WB; and the targeting relationship between miR-377-5p and HIF-1α gene was determined by Luciferase reporter gene assay.Results: The expression of miR-377-5p in HCC tissues was significantly lower than that in para-cancerous tissues (P<0.01). Compared with the control group, the expression of miR-377-5p in HepG2 cells of miR-377-5p mimic group elevated significantly, and the proliferation,invasion and the expression of N-caderin proteins decreased, significantly (all P<0.01), while the expression of E-caderin increased significantly (P<0.01). At the same time, the mRNA and protein expressions of HIF-1α in miR-377-5p mimic group decreased significantly (P<0.01 or P<0.05). miR-377-5p targetedly inhibited the expression of HIF-1α gene and suppressed the activation of VEGF pathway (all P<0.05). Conclusion: miR-377-5p inhibits the proliferation, invasion and EMT of HepG2 cells via targetedly inhibiting HIF-1α expression and suppressing the activation of VEGF signaling pathway.

四川省卫生和计划生育委员会科研资助项目（No.16PJ149）