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。
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.
作者
谷岩
张蕊
何菊
刘辉
Yan Gu;Rui Zhang;Ju He;Hui Liu(Department of Vascular Surgery,Tianjin First Central Hospital,Tianjin 300192,China)
出处
《中国现代医学杂志》
CAS
北大核心
2021年第23期38-44,共7页
China Journal of Modern Medicine
基金
天津市卫生局科技基金(No:2015KZ03)
天津市第一中心医院院内项目(No:CM201805)。