Longterm culture of telomerase-transduced rheumatoid arthritis fibroblast-like synoviocytes display a distinct gene expression pattern.
OBJECTIVE
To extend the lifespan of rheumatoid arthritis fibroblast-like synoviocytes (RA FLS) using human telomerase catalytic subunit (hTERT) and to test the hypothesis that longterm culture of hTERT-RA FLS may display a disease-specific gene expression pattern.


METHODS
RA 516 FLS were transduced by hTERT and the replicative properties of hTERT-RA 516 FLS were evaluated by repeated expansion. Gene expressions in hTERT-RA 516 FLS (passage 8) were compared with the gene expressions in hTERT-osteoarthritis (OA) 13A FLS (passage 8) by microarrays and RT-PCR. After continuous expansion in culture for an additional 4 months, gene expression was examined again using real-time RT-PCR and microarray.


RESULTS
While primary RA 516 FLS stopped dividing after repeated culture for about 120 days, hTERT-RA 516 FLS continued to grow at a steady rate. The hTERT-RA 156 FLS displayed a distinct gene expression pattern different from hTERT-OA 13A FLS. Several putative autoantigens and cytokine A2/monocyte chemoattractant protein-1 were expressed at significantly higher levels in longterm culture of hTERT-RA 516 FLS.


CONCLUSIONS
Telomerase-transduced RA FLS offer an alternative cell model for the study of RA and for examination of cellular/genetic alterations in RA FLS. Our findings provide further support for the notion that RA FLS are altered cells.
Sun
Yubo
Y
Department of Orthopaedic Surgery, Carolinas Medical Center, Charlotte, North Carolina,USA. yubo.sun@carolinashealthcare.org
Firestein
Gary
G
Boyle
David
D
Gruber
Helen
H
Cheung
Herman
H
D000328:Adult; D001172:Arthritis, Rheumatoid / Q000378:metabolism / Q000473:pathology; D015415:Biomarkers / Q000378:metabolism; D002478:Cells, Cultured; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D013582:Synovial Fluid / Q000166:cytology*; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic
D015415:Biomarkers; C400152:DNA enzyme ED5; D004277:DNA, Single-Stranded; D019098:Telomerase; D020781:Matrix Metalloproteinase 1