Objective: To determine whether miR-600 suppresses the proliferation of HeLa cells by inhibiting hypoxia-inducible factor-1α (HIF-1α) signaling pathway and its effect on expressions of cyclin D1 and vascular endothelial growth factor (VEGF).
Objective: HeLa cells were transfected with miR-600 mimic and plasmid-HIF-1α, either alone or in combination, to up-regulate miR-600 and HIF-1α expressions in the cells. Six hours after the transfection, the cell viability was assessed using MTT assay, and the mRNA and protein expressions of VEGF, cyclin D1, and HIF-1α were analyzed with qPCR and Western blotting.
Objective: The viability of HeLa cells showed no obvious changes 6 h after transfection with miR-600 mimic or Plasmid-HIF-1α. At 24 h and 48 h, the cells transfected with miR-600 mimic showed a time-dependent reduction of cell viability, while the cells transfected with Plasmid-HIF-1α alone and with both miR-600 mimic and Plasmid-HIF-1α showed increased cell viability. The cell viabilities in Plasmid-HIF-1α group were significantly higher than those in miR-600 mimic+Plasmid-HIF-1α group at 24 h and 48 h. Six hours after transfection with miR-600 mimic, the cells exhibited significantly decreased expressions of VEGF, cyclin D1, and HIF-1α, which were all significantly up-regulated in Plasmid-HIF-1α group and miR-600 mimic+Plasmid-HIF-1α group. VEGF, cyclin D1, and HIF-1α expressions were significant higher in Plasmid-HIF-1α group than in miR-600 mimic+ Plasmid-HIF-1α group.
Objective: miR-600 suppresses the proliferation of HeLa cells and down-regulate the expressions of cyclin D1 and VEGF by inhibiting HIF-1α signaling pathway.
目的: 探讨miR-600是否可通过HIF-1α信号通路调控宫颈癌HeLa细胞的增殖及对Cyclin D1和血管内皮生长因子（VEGF）表达的影响。
方法: 采用miR-600 mimic和Plasmid-HIF-1α转染HeLa细胞增加和诱导miR-600和HIF-1α的表达。qPCR和Western blot检测转染6 h后Plasmid-HIF-1α对于HeLa细胞HIF-1α表达的影响。将HeLa细胞分为空白对照组（无特殊处理）、miR-600 mimic组（转染miR-600拟似物miR-600 mimic）、Plasmid-HIF-1α组（P-HIF-1α组，转染Plasmid-HIF-1α）和miR-600 mimic+Plasmid-HIF-1α组（miR-600 mimic+P-HIF-1α组，同时转染miR-600 mimic和Plasmid-HIF-1α）。在转染6 h后，使用MTT法检测细胞活性，采用qPCR和Western blot测定VEGF、Cyclin D1和HIF-1α mRNA和蛋白的表达水平。
结果: 在转染6 h后，miR-600 mimic和Plasmid-HIF-1α对HeLa细胞活性无显著影响（P均 < 0.05）。但转染Plasmid-HIF-1α 6 h后，细胞HIF-1α表达水平显著增加。在转染24 h和48 h，与对照组相比较，miR-600 mimic组细胞活性呈时间依赖性下降，Plasmid-HIF-1α组和miR-600 mimic+Plasmid-HIF-1α组细胞活性呈时间依赖性增加，且Plasmid-HIF-1α组较miR-600 mimic+Plasmid-HIF-1α组细胞活性时间依赖性增加更加显著（P均 < 0.05）。转染6 h后，与对照组相比较，miR-600 mimic组VEGF、Cyclin D1和HIF-1α表达均明显下降，Plasmid-HIF-1α组和miR-600 mimic+Plasmid-HIF-1α组VEGF、Cyclin D1和HIF-1α表达均明显增加，且Plasmid-HIF-1α组较miR-600 mimic+Plasmid-HIF-1α组增加更明显（P均 < 0.05）。
结论: 在HeLa细胞中miR-600可以通过抑制HIF-1α信号通路下调Cyclin D1和VEGF的表达，从而抑制肿瘤细胞的增殖和分化。
Keywords: HeLa cells; cell proliferation; hypoxia-inducible factor-1α; miR-600; vascular endothelial growth factor.