Scleroderma fibroblasts demonstrate enhanced activation of Akt (protein kinase B) in situ.
Recent studies suggest that, in addition to activation and hypersecretion of matrix components, fibroblasts from patients with systemic sclerosis (SSc) are relatively resistant to apoptosis. Transforming growth factor-beta (TGF)-beta is strongly implicated in the pathogenesis of SSc and we and others have shown that TGF-beta can activate Akt, a kinase with potent anti-apoptotic effects. To determine whether Akt was activated in SSc, we quantified phospho-Akt expression in skin fibroblasts in vitro by western blot analysis and a functional kinase assay. In addition, the relative proportion of fibroblasts containing activated Akt in was quantified by immunohistochemistry on skin sections insitu. Analysis of Akt phosphorylation of skin fibroblasts in vitro suggested increased phosphorylation of Akt, and evaluation of skin sections by immunohistochemistry revealed significantly higher percentages of fibroblasts that stained for phospho-Akt compared with controls (78% +/- 14.0% vs 13% +/- 9%, p < 0.001). In addition, co-incident staining of phospho-Akt and alpha-smooth muscle actin was observed in some fibroblasts. These findings indicate that Akt is activated insitu in skin fibroblasts from patients with SSc. Akt activation may contribute to resistance to apoptosis, selection of disease-inducing fibroblasts, and, possibly, myofibroblasts.
Jun
Jae-Bum
JB
Division of Rheumatology, University of Washington School of Medicine, 1959 NE Pacific Street, Seattle 98195, Washington, USA.
Kuechle
Melanie
M
Min
Junki
J
Shim
Seung Cheol
SC
Kim
Giok
G
Montenegro
Vivianne
V
Korn
Joseph H
JH
Elkon
Keith B
KB
D017209:Apoptosis / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000201:enzymology* / Q000473:pathology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D012595:Scleroderma, Systemic / Q000378:metabolism* / Q000473:pathology*; D015398:Signal Transduction / Q000502:physiology; D012867:Skin / Q000201:enzymology / Q000473:pathology; D016212:Transforming Growth Factor beta / Q000378:metabolism
D011518:Proto-Oncogene Proteins; D016212:Transforming Growth Factor beta; C494918:AKT1 protein, human; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt