摘要
目的:深入研究微小RNA(miR)-30c靶向调控纤溶酶原激活物抑制物1(PAI-1)表达对血管内皮细胞活力和迁移能力的影响。方法:应用miR-30c模拟物(mimic)、抑制物(inhibitor)及阴性对照(NC)序列转染人脐静脉内皮细胞(HUVECs)后,RT-q PCR检测miR-30c水平及PAI-1的mRNA表达,Western blot法检测PAI-1蛋白的表达,CCK-8法和划痕实验分别检测细胞活力和迁移能力。生物信息学方法预测miR-30c与PAI-1的mRNA 3’-UTR结合位点,并用双萤光素酶报告基因验证miR-30c对PAI-1 mRNA的靶向作用。结果:miR-30c能够靶向调控PAI-1的mRNA和蛋白表达,与对照组和NC序列转染组比较,增加miR-30c表达可导致PAI-1的mRNA和蛋白表达降低,进而增强HUVECs的活力和迁移能力;相反,抑制miR-30c表达可导致PAI-1的mRNA和蛋白表达升高,进而抑制HUVECs的活力和迁移能力。结论:高表达miR-30c可抑制PAI-1表达,导致HUVECs活力和迁移能力明显增强,提示miR-30c可能参与调节内皮细胞功能。
AIM：To investigate the effect of microRNA（ miR）-30 c on the viability and migratory ability of human umbilical vein endothelial cells（ HUVECs） by targeting plasminogen activator inhibitor-1（ PAI-1）.METHODS：The HUVECs were transfected with miR-30 c mimic and inhibitor or negative control（ NC）,and then the expression levels of miR-30 c,PAI-1 mRNA and protein were detected by RT-q PCR and Western blot.The viability and migratory ability of HUVECs were measured by CCK-8 assay and wound healing test.After bioinformatic analysis,the assessment of miR-30 c binding to PAI-1 3＇-UTR was carried out using a luciferase reporter gene assay.RESULTS：miR-30 c directly down-regulated PAI-1 levels by binding to the 3＇UTR seed sequence of PAI-1 mRNA.Furthermore,transfection of a miR-30 c mimic down-regulated the expression of PAI-1 at mRNA and protein levels,leading to enhanced migratory ability and viability of the HUVECs.However,transfection of a miR-30 c inhibitor up-regulated the expression of PAI-1 at mRNA and protein levels,leading to decreased migratory ability and viability.CONCLUSION：Regulation of miR-30 c level changes the migratory ability and viability of HUVECs by affecting the PAI-1 expression,indicating the involvement of miR-30 c in modulating endothelial function.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2016年第12期2199-2204,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81172050
No.81570263)
四川省科技厅课题(No.2014FZ0104)
四川省教育厅课题(No.16ZA0178)
作者简介
通讯作者Tel：0830-3161673;E-mail：jbwucn@163.com
