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
Viral microRNAs are one component of the RNA interference phenomenon generated during viral infection. They were first identified in the Herpesviridae family, where they were found to regulate viral mRNA translation. ...Viral microRNAs are one component of the RNA interference phenomenon generated during viral infection. They were first identified in the Herpesviridae family, where they were found to regulate viral mRNA translation. In addition, prior work has suggested that Kaposi's sarcoma-associated herpesvirus (KSHV) is capable of regulating cellular gene transcription by miRNA. We demonstrate that a miRNA, hsv1-mir-H27, encoded within the genome of herpes simplex virus 1 (HSV-1), targets the mRNA of the cellular transcriptional repressor Kelch-like 24 (KLHL24) that inhibits transcriptional efficiency of viral immediate-early and early genes. The viral miRNA is able to block the expression of KLHL24 in cells infected by HSV-1. Our discovery reveals an effective viral strategy for evading host cell defenses and supporting the efficient replication and proliferation of HSV-1.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.
基金supported by the National Natural Science Foundation of China (30670094, 30700028)National Basic Research Program of China (2012CB518901, 2011CB504903)
文摘Viral microRNAs are one component of the RNA interference phenomenon generated during viral infection. They were first identified in the Herpesviridae family, where they were found to regulate viral mRNA translation. In addition, prior work has suggested that Kaposi's sarcoma-associated herpesvirus (KSHV) is capable of regulating cellular gene transcription by miRNA. We demonstrate that a miRNA, hsv1-mir-H27, encoded within the genome of herpes simplex virus 1 (HSV-1), targets the mRNA of the cellular transcriptional repressor Kelch-like 24 (KLHL24) that inhibits transcriptional efficiency of viral immediate-early and early genes. The viral miRNA is able to block the expression of KLHL24 in cells infected by HSV-1. Our discovery reveals an effective viral strategy for evading host cell defenses and supporting the efficient replication and proliferation of HSV-1.
