Zhou X, Deng J, Zhang W, Wang J
Department of Obstetrics and Gynecology, Chengdu Second People's Hospital, Chengdu 610000, China.
Department of Obstetrics and Gynecology, First Affiliated Hospital of Chengdu Medical College, Chengdu 610000, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2021 Feb 25;41(2):210-215. doi: 10.12122/j.issn.1673-4254.2021.02.07.
To determine whether miR-600 suppresses the proliferation of HeLa cells by inhibiting hypoxia-inducible factor-1 (HIF-1) signaling pathway and its effect on expressions of cyclin D1 and vascular endothelial growth factor (VEGF).
HeLa cells were transfected with miR-600 mimic and plasmid-HIF-1, either alone or in combination, to up-regulate miR-600 and HIF-1 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-1 were analyzed with qPCR and Western blotting.
The viability of HeLa cells showed no obvious changes 6 h after transfection with miR-600 mimic or Plasmid-HIF-1. 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-1 alone and with both miR-600 mimic and Plasmid-HIF-1 showed increased cell viability. The cell viabilities in Plasmid-HIF-1 group were significantly higher than those in miR-600 mimic+Plasmid-HIF-1 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-1, which were all significantly up-regulated in Plasmid-HIF-1 group and miR-600 mimic+Plasmid-HIF-1 group. VEGF, cyclin D1, and HIF-1 expressions were significant higher in Plasmid-HIF-1 group than in miR-600 mimic+ Plasmid-HIF-1 group.
miR-600 suppresses the proliferation of HeLa cells and down-regulate the expressions of cyclin D1 and VEGF by inhibiting HIF-1 signaling pathway.
确定miR-600是否通过抑制缺氧诱导因子-1（HIF-1）信号通路来抑制HeLa细胞增殖及其对细胞周期蛋白D1和血管内皮生长因子（VEGF）表达的影响。
将miR-600模拟物和质粒-HIF-1单独或联合转染HeLa细胞，以上调细胞中miR-600和HIF-1的表达。转染6小时后，使用MTT法评估细胞活力，并用qPCR和蛋白质印迹法分析VEGF、细胞周期蛋白D1和HIF-1的mRNA和蛋白质表达。
用miR-600模拟物或质粒-HIF-1转染HeLa细胞6小时后，细胞活力无明显变化。在24小时和48小时时，转染miR-600模拟物的细胞显示出细胞活力呈时间依赖性降低，而单独转染质粒-HIF-1以及同时转染miR-600模拟物和质粒-HIF-1的细胞显示细胞活力增加。在24小时和48小时时，质粒-HIF-1组的细胞活力显著高于miR-600模拟物+质粒-HIF-1组。用miR-600模拟物转染6小时后，细胞中VEGF、细胞周期蛋白D1和HIF-1的表达显著降低，而在质粒-HIF-1组和miR-600模拟物+质粒-HIF-1组中这些表达均显著上调。质粒-HIF-1组中VEGF、细胞周期蛋白D1和HIF-1的表达显著高于miR-600模拟物+质粒-HIF-1组。
miR-600通过抑制HIF-1信号通路抑制HeLa细胞增殖并下调细胞周期蛋白D1和VEGF的表达。
