目的:为进一步探讨miRNA-24是否参与内皮型一氧化氮合酶(eNOS)表达的调控及其对血管内皮细胞增殖的影响,本研究构建miRNA-24高表达质粒,并用脂质体将其导入人脐静脉内皮细胞(HUVECs),观察miRNA-24对eNOS表达及HUVECs增殖的影响。方法:...目的:为进一步探讨miRNA-24是否参与内皮型一氧化氮合酶(eNOS)表达的调控及其对血管内皮细胞增殖的影响,本研究构建miRNA-24高表达质粒,并用脂质体将其导入人脐静脉内皮细胞(HUVECs),观察miRNA-24对eNOS表达及HUVECs增殖的影响。方法:用四甲基偶氮唑蓝(MTT)法检测细胞增殖情况,RT-PCR、免疫组织化学和Western blotting检测eNOS与Sp1转录因子mRNA和蛋白表达水平。结果:与对照组比较,miRNA-24高表达组细胞增殖减慢41.97%(0.47±0.04 vs 0.81±0.03,P<0.01),eNOS mRNA降低44.8%(0.48±0.01 vs 0.87±0.03,P<0.05),蛋白表达减少71.92%(0.16±0.06 vs 0.57±0.08,P<0.05);同时Sp1 mRNA降低53.00%(0.45±0.02 vs 0.93±0.01,P<0.05),其蛋白质表达量也相应减少62.31%(0.13±0.07 vs 0.31±0.09,P<0.05)。miRNA-24抑制组中,上述指标比对照组降低,但比miRNA-24高表达组明显升高。结论:miRNA-24高表达明显抑制HUVECs的增殖及eNOS的表达;Sp1可能是参与miRNA-24调控eNOS表达的重要因素之一。展开更多
Objective:To investigate the effect and mechanism of miRNA-24 on the proliferation,migration and tube formation of vascular endothelial cells.Methods:Human umbilical vein endothelial cells(HUVECs)were assigned into co...Objective:To investigate the effect and mechanism of miRNA-24 on the proliferation,migration and tube formation of vascular endothelial cells.Methods:Human umbilical vein endothelial cells(HUVECs)were assigned into control,microRNA(miR)-24 overexpression and anti-miR-24 groups.The proliferation and migration abilities of HUVECs were detected by methyl thiazolyl tetrazolium(MTT)assay and scratch wound healing assay,respectively.The ability of HUVECs to form tubular structures was evaluated by a tube formation assay.The mRNA and protein expressions of vascular endothelial growth factor(VEGF)and transcription factor Sp1 were determined by RT-PCR,immunocytochemistry and western blotting,respectively.Results:The miR-24 overexpression group exhibited decreased cell proliferation and migration,and expressions of VEGF and Sp1 compared with the control group(P <0.01).No tube-like network structure was formed in the miR-24 overexpression group.However,inhibition of miR-24 in HUVECs markedly increased cell proliferation and migration,enhanced tube formation and expressions of VEGF and Sp1(P<0.05 or P<0.01).Conclusion:MiR-24 suppressed the proliferation,migration and tube formation of HUVECs,and the mechanism might be related to the down-regulation of VEGF expression.Sp1 might participate in this regulation process.展开更多
文摘目的:为进一步探讨miRNA-24是否参与内皮型一氧化氮合酶(eNOS)表达的调控及其对血管内皮细胞增殖的影响,本研究构建miRNA-24高表达质粒,并用脂质体将其导入人脐静脉内皮细胞(HUVECs),观察miRNA-24对eNOS表达及HUVECs增殖的影响。方法:用四甲基偶氮唑蓝(MTT)法检测细胞增殖情况,RT-PCR、免疫组织化学和Western blotting检测eNOS与Sp1转录因子mRNA和蛋白表达水平。结果:与对照组比较,miRNA-24高表达组细胞增殖减慢41.97%(0.47±0.04 vs 0.81±0.03,P<0.01),eNOS mRNA降低44.8%(0.48±0.01 vs 0.87±0.03,P<0.05),蛋白表达减少71.92%(0.16±0.06 vs 0.57±0.08,P<0.05);同时Sp1 mRNA降低53.00%(0.45±0.02 vs 0.93±0.01,P<0.05),其蛋白质表达量也相应减少62.31%(0.13±0.07 vs 0.31±0.09,P<0.05)。miRNA-24抑制组中,上述指标比对照组降低,但比miRNA-24高表达组明显升高。结论:miRNA-24高表达明显抑制HUVECs的增殖及eNOS的表达;Sp1可能是参与miRNA-24调控eNOS表达的重要因素之一。
基金supported by the National Natural Science Foundation of China(No.81373403)
文摘Objective:To investigate the effect and mechanism of miRNA-24 on the proliferation,migration and tube formation of vascular endothelial cells.Methods:Human umbilical vein endothelial cells(HUVECs)were assigned into control,microRNA(miR)-24 overexpression and anti-miR-24 groups.The proliferation and migration abilities of HUVECs were detected by methyl thiazolyl tetrazolium(MTT)assay and scratch wound healing assay,respectively.The ability of HUVECs to form tubular structures was evaluated by a tube formation assay.The mRNA and protein expressions of vascular endothelial growth factor(VEGF)and transcription factor Sp1 were determined by RT-PCR,immunocytochemistry and western blotting,respectively.Results:The miR-24 overexpression group exhibited decreased cell proliferation and migration,and expressions of VEGF and Sp1 compared with the control group(P <0.01).No tube-like network structure was formed in the miR-24 overexpression group.However,inhibition of miR-24 in HUVECs markedly increased cell proliferation and migration,enhanced tube formation and expressions of VEGF and Sp1(P<0.05 or P<0.01).Conclusion:MiR-24 suppressed the proliferation,migration and tube formation of HUVECs,and the mechanism might be related to the down-regulation of VEGF expression.Sp1 might participate in this regulation process.
