Detection of oncofetal h19 RNA in rheumatoid arthritis synovial tissue.
The expression of oncofetal H19 RNA and its localization/cellular source was analyzed in synovial tissue (ST) and isolated synovial macrophages (Mphi) or synovial fibroblasts (SFBs) by reverse transcriptase-polymerase chain reaction (RT-PCR), in situ hybridization, and immunohistochemistry. RT-PCR showed significantly higher H19 expression in ST from patients with rheumatoid arthritis (RA) (P = 0.000) and osteoarthritis (OA) (P = 0.009) than in normal/joint trauma controls (N/JT), but comparable levels in reactive arthritis. In situ hybridization demonstrated strong signals in all RA-ST samples (n = 8), with > or =85% positive cells in the lining layer, diffuse infiltrates, and stroma regions. In lymphoid aggregates and endothelial cells only 20% were positive. RA-ST contained a significantly higher percentage of strongly positive lining cells than OA-ST and N/JT-ST. H19 RNA was expressed in both Mphi and SFBs, as confirmed by RT-PCR in isolated RA Mphi and SFBs (n = 3). In RA-SFBs, low constitutive H19 RNA expression in culture (10% fetal calf serum) was strongly increased on starvation (3.5-fold, 1% fetal calf serum), with or without the addition of interleukin-1beta (10 to 100 U/ml), tumor necrosis factor-alpha (1 to 25 ng/ml), or platelet-derived growth factor-BB (2.5 to 10 U/ml). In OA-SFBs, this starvation-induced increase was lower (twofold), reaching significant differences compared with RA-SFBs after stimulation with interleukin-1beta and platelet-derived growth factor-BB. In both RA- and OA-SFBs, the MAP-kinase ERK-1/2 pathway and the phosphatidylinositol-3 kinase pathway influenced H19 RNA expression, as shown by inhibitor studies. Significant overexpression of H19 RNA and its increased sensitivity to starvation/cytokine regulation in RA suggests a pathogenetic role of this oncofetal gene, possibly reflecting embryonal dedifferentiation of the adult ST and/or ongoing inflammatory/oxidative stress.
Stuhlmüller
Bruno
B
Department of Internal Medicine, Rheumatology and Clinical Immunology, Charité University Hospital, Humboldt University of Berlin, Tucholskystrasse 2, D-10117 Berlin, Germany. bruno.stuhlmuller@charite.de
Kunisch
Elke
E
Franz
Juliane
J
Martinez-Gamboa
Lorena
L
Hernandez
Maria M
MM
Pruss
Axel
A
Ulbrich
Norbert
N
Erdmann
Volker A
VA
Burmester
Gerd R
GR
Kinne
Raimund W
RW
D000328:Adult; D000368:Aged; D000951:Antigens, Neoplasm / Q000378:metabolism*; D016918:Arthritis, Reactive / Q000378:metabolism; D001172:Arthritis, Rheumatoid / Q000378:metabolism*; D000077214:Becaplermin; D016022:Case-Control Studies; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D007375:Interleukin-1 / Q000494:pharmacology; D007596:Joints / Q000293:injuries; D008264:Macrophages / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D010003:Osteoarthritis / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D020574:Proto-Oncogene Proteins c-sis; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013583:Synovial Membrane / Q000378:metabolism* / Q000473:pathology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014947:Wounds and Injuries / Q000378:metabolism
D000951:Antigens, Neoplasm; D003470:Culture Media; C094335:H19 long non-coding RNA; D007375:Interleukin-1; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D014409:Tumor Necrosis Factor-alpha; C031192:oncofetal antigens; D000077214:Becaplermin