Regulation of synoviocyte proliferation, apoptosis, and invasion by the p53 tumor suppressor gene.
Recent studies show that 1) the p53 tumor suppressor protein is overexpressed by rheumatoid arthritis (RA) synovium and fibroblast-like synoviocytes (FLS) and 2) somatic mutations previously identified in human tumors are present in RA synovium and FLS. We have hypothesized that abnormalities in p53 can contribute to chronic destructive RA synovitis. To understand the functional consequences of p53 abnormalities in FLS, RA and normal FLS expressing wild-type p53 were transduced with a retroviral vector encoding the human papilloma virus 18 E6 gene, which inactivates endogenous p53 protein. Three RA and one normal FLS lines were infected with recombinant retrovirus encoding the neomycin resistance gene (neo) or E6+neo. FLS proliferation, apoptosis, and invasion was studied in E6, neo, and uninfected parental strains (PS). The growth rate for E6 was significantly increased with a sixfold increase in cell number after 7 days compared with a twofold to threefold increase in neo and PS. When FLS were treated with cytokines, proliferative response of E6, neo, and PS to interleukin-1 and transforming growth factor-beta were similar. However, response to platelet-derived growth factor was significantly greater in E6 FLS compared with neo or PS. Apoptosis was studied by incubating FLS with sodium nitroprusside as a source of nitric oxide or hydrogen peroxide for 8 hours and examining DNA fragmentation and E6 cells were significantly less susceptible to cell death. In addition, E6 FLS were more invasive into cartilage extracts than neo or PS using an in vitro cell invasion assay. These data suggest that p53 is a critical regulator of FLS proliferation, apoptosis, and invasiveness. Abnormalities of p53 function might contribute to synovial lining expansion and joint destruction in RA.
Aupperle
K R
KR
University of California, San Diego, School of Medicine, La Jolla 92093-0656, USA.
Boyle
D L
DL
Hendrix
M
M
Seftor
E A
EA
Zvaifler
N J
NJ
Barbosa
M
M
Firestein
G S
GS
D017209:Apoptosis / Q000235:genetics*; D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000473:pathology*; D015152:Blotting, Northern; D002455:Cell Division / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D002478:Cells, Cultured; D017364:Collagenases / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D009856:Oncogene Proteins, Viral / Q000235:genetics; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D012097:Repressor Proteins / Q000235:genetics; D013583:Synovial Membrane / Q000473:pathology*; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*
C068651:E6 protein, Human papillomavirus type 16; D007375:Interleukin-1; D009856:Oncogene Proteins, Viral; D010982:Platelet-Derived Growth Factor; D012097:Repressor Proteins; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D016159:Tumor Suppressor Protein p53; D009599:Nitroprusside; D006861:Hydrogen Peroxide; D017364:Collagenases