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目的: 分析肝癌组织缺氧诱导因子-1α(HIF-1α)表达特征及沉默HIF-1 α对癌细胞增殖的影响。方法:以免疫组化法分析自身配对的肝癌及癌周组织中HIF-1α表达。HepG2细胞转染靶向HIF-1 α的miRNA干扰质粒,RT-PCR、Western blot检测HIF-1α基因及蛋白水平变化;酶联免疫吸附法检测培养液血管内皮生长因子(VEGF)、血管生成素(ANG)-2表达;阿霉素预处理后,以Annexin V-FLUOS/PI双染法分析HepG2细胞凋亡率,并检测细胞周期改变。结果:肝癌及周围组织HIF-1α呈棕黄色颗粒状表达,定位于胞浆和胞核,其特征是癌旁组织全数表达高于癌灶组织(80.0%,χ~2=22.35,P<0.001),癌灶组织HIF-1α表达与肿瘤大小相关。HepG2细胞转染靶向HIF-1αmiRNA干扰质粒后72 h,HIF-1α在转录水平下降87%,蛋白水平下降56%;培养液中VEGF和ANG-2表达水平分别减少54%和36%;HepG2细胞凋亡率为(22.46±0.61)%,G_1期和S期比率分别为(61.49±1.12)%和(22.40±0.58)%;联合阿霉素后HepC2细胞凋亡率为(36.99±0.88)%,G_1期和S期比率分别为(65.68±0.91)%和(19.47±1.34)%。结论:肝癌组织HIF-1α过表达与肝癌发展相关,沉默HIF-1 α可有效调控下游血管生成相关基因表达,抑制癌细胞增殖,改善化疗药物敏感性
Objective: To investigate the expression of hypoxia inducible factor-1α (HIF-1α) in hepatocellular carcinoma and the effect of silencing HIF-1α on the proliferation of cancer cells. Methods: The expression of HIF-1α in paired hepatocellular carcinoma and its adjacent tissues was analyzed by immunohistochemistry. HepG2 cells transfected with miRNA interference targeting HIF-1α plasmid, RT-PCR, Western blot detection of HIF-1αgene and protein levels; enzyme-linked immunosorbent assay vascular endothelial growth factor (VEGF), angiogenin (ANG) -2. After pretreatment with doxorubicin, the apoptosis rate of HepG2 cells was analyzed by Annexin V-FLUOS / PI double staining and the cell cycle was detected. Results: The expression of HIF-1α in hepatocellular carcinoma and its surrounding tissues was buffy granules and located in the cytoplasm and nucleus. HIF-1α expression in hepatocellular carcinoma and its surrounding tissues was higher than that in carcinoma tissues (80.0%, χ ~ 2 = 22.35, P <0.001) ), The expression of HIF-1α in tumor tissue correlated with tumor size. HIF-1α decreased by 87% and protein level by 56% at 72 h after transfection with HepG2 cells targeting HIF-1αmiRNA. The expression levels of VEGF and ANG-2 decreased by 54% and 36% respectively in HepG2 cells. HepG2 The apoptotic rate of HepC2 cells was (22.46 ± 0.61)%, and the rates of G_1 phase and S phase were (61.49 ± 1.12)% and (22.40 ± 0.58)%, respectively. %, G_1 and S rates were (65.68 ± 0.91)% and (19.47 ± 1.34)%, respectively. CONCLUSIONS: HIF-1α overexpression is associated with the development of liver cancer in HCC. Silencing HIF-1α can effectively regulate the expression of downstream angiogenesis-related genes, inhibit the proliferation of cancer cells, and improve the chemosensitivity