MRTF-A的SUMO化对人脐静脉内皮细胞迁移能力的影响

Inhibition of CCN1 expression by sumoylation of MRTF-A on migration of human umbilical vein endothelial cells

摘要:

目的 探讨MRTF-A的SUMO化抑制CCN1的表达对人脐静脉内皮细胞（HUVECs）迁移能力的影响。方法 采用实时荧光定量聚合酶链反应（qRT-PCR）和Western blotting检测CCN1的表达。通过免疫共沉淀（co-IP）和染色质免疫沉淀（ChIP）分析MRTF-A与小泛素相关修饰物（SUMO）的相互作用。采用细胞划痕实验和Transwell法检测细胞迁移能力。结果 CCN1的转录活性受活化的PIAS1、UBC9或SUMO1抑制。SUMOylation位点的突变抑制MRTF-A对CCN1的转录活性。细胞迁移实验表明，MRTF-A的SUMO化抑制HUVECs的迁移（P <0.05）。结论 MRTF-A的SUMO化通过靶向CCN1抑制HUVECs的迁移。

Abstract:

Objective To investigate whether MRTF-A SUMOylation inhibited CCN1 expression in human umbilical vein endothelial cells (HUVECs).Methods CCN1 expression was detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting analysis. MRTF-A and small ubiquitin-like modifier 1 (SUMO1) interactions were analysed by co-immunoprecipitation and chromosomal immunoprecipitation. Cell migration capacity was detected using cell migration and Transwell chamber assays.Results It was found that CCN1 transcriptional activity was inhibited through a mechanism dependent on attenuation of the SUMO modifications of MRTF-A by SUMO1/protein inhibitor of activated STAT 1 or UBC9. Moreover, mutation of the SUMOylation sites enhanced the transcriptional activity of MRTF-A, and abolished the repression of CCN1. Cell migration assays showed that MRTF-A SUMOylation inhibited the migration of HUVECs (P < 0.05).Conclusions These results suggest that MRTF-A SUMOylation inhibited the migration of HUVECs by targeting CCN1.