miR-29b调控HBP1信号通路与RA患者PBMs的关系

miR-29b regulates HBP1 signaling pathway and PBMs in RA patients

目的:探讨miR-29b调控高迁移率族(HMG)转录因子1(HBP1)与类风湿关节炎(RA)患者CD14阳性外周血单核细胞(PBMs)的关系。方法:使用Target Scan和miRDB数据库鉴定miR-29b的直接靶点HBP1。使用ViaFect试剂建立稳定表达外源性HBP1的THP-1细胞(THP-1/HBP1),将miR-29b模拟物(miR-29b mimics)或模拟物阴性对照(Mimics-NC)进行转染,检测细胞存活和凋亡情况。采用RT-qPCR法检测靶基因的表达水平。采用荧光素酶报告试剂盒检测报告基因。采用Western blotting检测HBP1的表达。结果:使用Target Scan和miRDB数据库筛选出20个可能是miR-29b潜在的靶点基因,其中HBP1是一种与凋亡相关的转录因子。在RA PBMs中,HBP1表达水平与miR-29b的表达水平具有负相关性;miR-29b可以直接与HBP1的3'-UTR结合,并抑制其在PBMs中的转录。结论:RA患者PBMs的miR-29b调控HBP1信号通路可能是RA的发病机制之一。

Objective:To investigate the relationship between miR-29b regulation of high mobility group(HMG)transcription factor 1(HBP1)and CD14-positive peripheral blood mononuclear cells(PBMs)in patients with rheumatoid arthritis(RA).Methods:Target Scan and miRDB database were used to identify HBP1,the direct target of miR-29b.ViaFect was used to establish THP-1 cells(THP-1/HBP1)that stably expressed exogenous HBP1.miR-29b mimics(miR-29b mimics)or mimics negative control(MIMicS-NC)were transfected to detect cell survival and apoptosis.The expression level of target genes was detected by RT-qPCR.Luciferase reporter kit was used to detect reporter genes.The expression of HBP1 was detected by Western blotting.Results:Twenty potential Target genes of miR-29b were screened using Target Scan and miRDB database,among which HBP1 was a transcription factor associated with apoptosis.In RA PBMs,the expression level of HBP1 was negatively correlated with the expression level of miR-29b.miR-29b can directly bind to the 3’-UTR of HBP1 and inhibit its transcription in PBMs.Conclusion:The regulation of HBP1 signaling pathway by miR-29b of PBMs in RA patients may be one of the pathogenesis o f RA.