miR-21-3p靶向PTEN对深静脉血栓内皮祖细胞的增殖和凋亡的影响

Effect of miR-21-3p Targeting PTEN on the Proliferation and Apoptosis of Endothelial Progenitor Cells in Deep Vein Thrombosis

目的探讨miR-21-3p对深静脉血栓(deep vein thrombosis,DVT)中内皮祖细胞增殖和凋亡的影响及其作用机制。方法采用RT-PCR检测DVT患者和健康受试者外周血中miR-21-3p和PTEN的表达水平;在DVT内皮祖细胞中转染miR-21-3p mimics和miR-21-3p inhibitors后,采用CCK8法检测细胞增殖情况,流式细胞仪检测细胞凋亡情况,RTPCR和western blot检测PTEN mRNA和蛋白表达水平。双荧光素酶报告检测miR-21-3p和PTEN的结合情况。结果 miR-21-3p在DVT患者外周血中的表达水平明显低于健康受试者(P<0.05),同时miR-21-3p和PTEN在DVT患者外周血中的表达水平呈明显负相关(P<0.05)。在DVT内皮祖细胞中转染miR-21-3p mimics后,细胞增殖能力较miR-NC组明显升高(P<0.05),细胞凋亡率较miR-NC组明显降低(P<0.05)。在DVT内皮祖细胞中转染miR-21-3p inhibitors后,细胞增殖能力较miR-NC组明显降低(P<0.05),细胞凋亡率较miR-NC组明显升高(P<0.05)。双荧光素酶结果显示miR-21-3p能够靶向结合PTEN。在DVT内皮祖细胞中共转染miR-21-3p mimics和LV-PTEN后能够反转单独转染miR-21-3p mimics对细胞增殖和凋亡的影响(P均>0.05)。结论 miR-21-3p在DVT中明显低表达,过表达miR-21-3p能够通过靶向调控PTEN促进内皮祖细胞增殖和抑制细胞凋亡。miR-21-3p可能成为DVT治疗的一个潜在作用靶点。

Objective To investigate the effect of miR-21-3p on proliferation and apoptosis of endothelial progenitor cells,and explore the underlying mechanism in deep vein thrombosis(DVT).Methods The expression level of miR-21-3p and PTEN in peripheral blood of DVT patients and healthy subjects were analyzed by real-time polymerase chain reaction(RT-PCR).After DVT endothelial progenitor cells transfected with miR-21-3p mimics and inhibitors,the cell proliferation was detected by CCK8 assay,the apoptosis rate was measured by Flow cytometry analysis,and the expression level of PTEN was detected by RT-PCR and western blot.The binding of miR-21-3p and PTEN was detected by double luciferase assay.Results The expression level of miR-21-3p in peripheral blood of DVT patients was significantly lower than that of healthy subjects(P<0.05).Meanwhile,the expression levels of miR-21-3p and PTEN in peripheral blood of patients with DVT were significantly negatively correlated(P<0.05).After DVT endothelial progenitor cell transfected with miR-21-3p mimics,cell proliferation ability was significantly increased compared with miR-NC group(P<0.05),and the apoptosis rate was significantly decreased compared with miR-NC group(P<0.05).After DVT endothelial progenitor cell transfected with miR-21-3p inhibitors,cell proliferation ability was significantly decreased compared with miR-NC group(P<0.05),and the apoptosis rate was significantly increased compared with miR-NC group(P<0.05).Double luciferase results showed that miR-21-3p could bind PTEN in a targeted manner.After co-transfection with miR-21-3p mimics and LV-PTEN in DVT endothelial progenitor cells,the effects of miR-21-3p mimics on cell proliferation and apoptosis were reversed(P>0.05).Conclusion MiR-21-3p is significantly down-regulated in DVT.Over-expression of miR-21-3p can promote the proliferation of endothelial progenitor cells and inhibit apoptosis through targeting PTEN.MiR-21-3p may be a potential target for DVT treatment.