title: [Activation of tumor necrosis factor receptor-associated factor 6 in anti-β2GPI/β2GPI-induced tissue factor expression on THP-1 cells].
abstract: OBJECTIVE
  To investigate whether tumor necrosis factor receptor-associated factor 6 (TRAF6) is involved in anti-β2GPI/β2GPI-induced tissue factor (TF) expression on THP-1 cells.
  
  
  METHODS
  The total RNA was extracted and the protein lysates were collected from THP-1 cells stimulated with anti-β2GPI/β2GPI complex. And then the TF expression on THP-1 cells was detected by real-time quatitative PCR (RT-qPCR) and TF activity kit. TRAF6 mRNA and its protein expression were investigated by RT-qPCR and Western blotting, respectively. The proteasome inhibitor, MG-132, was used for inhibitory assays, in order to demonstrate the effect of anti-β2GPI/β2GPI complex on THP-1 cells.
  
  
  RESULTS
  The TF expression (both mRNA and activity) on THP-1 cells was significantly up-regulated with the treatment of anti-β2GPI/β2GPI complex (100 mg/L), compared with untreated cells(P<0.05). The TRAF6 mRNA and protein levels in THP-1 cells were also significantly increased with the treatment of anti-β2GPI/β2GPI complex. The expression of TRAF6 was shown in a time-dependent manner, with the maximal level at 15 minutes (mRNA) and 30 minutes (protein) respectively. All the stimulating effects of anti-β2GPI/β2GPI complex (100 mg/L) on THP-1 cells were inhibited by MG-132 (5 μmol/L).
  
  
  CONCLUSIONS
  TRAF6 is up-regulated and contributes to TF expression on THP-1 cells induced with anti-β2GPI/β2GPI complex.
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212013, China. peakxuguoying@126.com
authors.forename: Guo-ying
authors.initials: GY
authors.lastname: Xu
authors.forename: Hong
authors.initials: H
authors.lastname: Zhou
authors.forename: Hai-ping
authors.initials: HP
authors.lastname: Wen
authors.forename: Dong-lin
authors.initials: DL
authors.lastname: Guo
authors.forename: Fang
authors.initials: F
authors.lastname: Zhou
authors.forename: Dong-dong
authors.initials: DD
authors.lastname: Chen
authors.forename: Hong-xiang
authors.initials: HX
authors.lastname: Xie
mesh_terms: D017152:Antibodies, Antiphospholipid / Q000276:immunology / Q000494:pharmacology*; D002478:Cells, Cultured; D006801:Humans; D048029:TNF Receptor-Associated Factor 6 / Q000235:genetics / Q000378:metabolism*; D013925:Thromboplastin / Q000096:biosynthesis* / Q000235:genetics; D015854:Up-Regulation; D053482:beta 2-Glycoprotein I / Q000276:immunology / Q000494:pharmacology*
chemical_list: D017152:Antibodies, Antiphospholipid; D048029:TNF Receptor-Associated Factor 6; D053482:beta 2-Glycoprotein I; D013925:Thromboplastin