Skip to main content
  • Meeting abstract
  • Published:

AKT regulates TNF-alpha-mediated apoptosis of rheumatoid arthritis synovial fibroblasts


To determine whether TNF-α driven proliferation of rheumatoid synovial fibroblasts is associated with upregulation of the serine/threonine kinase B (PKB)/AKT activity and RA synovial fibroblast survival.


Staining of phosphorylated AKT was done using antiphosphorylated Thr308-AKT antibody. Phosphorylated AKT was analyzed by Western blot, and AKT activity was analyzed using a kinase assay. The cytotoxicity of TNF-α treatment or TNF-α plus AKT activity inhibitor wortmannin, TNF-α plus dominant mutant (AdAkt-DN) or AdPTEN was analyzed using ATPLite assay.


The levels of phosphorylated-Akt are higher in RASF than in OASF, as demonstrated by immunohistochemical staining, immunoblot analysis and an Akt kinase assay. The levels of phosphorylated Akt and Akt kinase activity were increased by stimulation of primary RASF with TNF-α (10 ng/ml). Treatment of RASF with the PI 3-kinase inhibitor, wortmannin (50 nM), plus TNF-α resulted in apoptosis of 75 ± 8% of RASF within 24 h. This proapoptosis effect was specific for Akt, as equivalent levels of apoptosis were observed upon TNF-α treatment of RASF trans-fected with adenovirus expressing a dominant negative-Akt (AdAkt-DN) and with an adenovirus expressing PTEN (AdPTEN), which opposes the action of Akt.


These results indicate that phosphorylated Akt acts as a survival signal in RASF and contributes to the stimulatory effect of TNF-α on these cells by inhibiting the apoptosis response. This effect was not observed in OSAF, and may reflect the pathophysiologic changes associated with the proliferating synovium in rheumatoid arthritis.

Author information

Authors and Affiliations


Rights and permissions

Reprints and permissions

About this article

Cite this article

Mountz, J., Zhang, HG., Wang, Y. et al. AKT regulates TNF-alpha-mediated apoptosis of rheumatoid arthritis synovial fibroblasts. Arthritis Res Ther 3 (Suppl 1), P8 (2001).

Download citation

  • Received:

  • Published:

  • DOI: