Meyringer, Rotraud and Neumann, Elena and Judex, Martin and Landthaler, Michael and Kullmann, Frank and Schoelmerich, Jorgen and Gay, Steffen and Tarner, Ingo H. and Distler, Oliver and Mueller-Ladner, Ulf (2007) Analysis of gene expression patterns in systemic sclerosis fibroblasts using RNA arbitrarily primed-polymerase chain reaction for differential display. JOURNAL OF RHEUMATOLOGY, 34 (4). pp. 747-753. ISSN 0315-162X, 1499-2752
Full text not available from this repository. (Request a copy)Abstract
Objective. To identify genes that are differentially expressed in systemic sclerosis (SSc) fibroblasts of clinically involved and noninvolved skin compared to normal dermal fibroblasts, using RNA arbitrarily primed-polymerase chain reaction (RAP-PCR) for differential display. Methods. We examined 12 fibroblast cultures derived from clinically involved skin, 3 fibroblast cultures from noninvolved skin. and 4 fibroblast cultures from healthy skin. After extraction of total RNA, the first step of RAP-PCR was performed using different arbitrary 10-12-base primers for first-strand cDNA synthesis. Second-strand synthesis was achieved by cycling using different arbitrary 10-base primers, followed by sequence analysis of the amplified fingerprint products. The resulting sequences were aligned to the GenBank (R) database using Blast (R) Search. Confirmation of differential expression was performed with specific primers using real-time PCR. Results. Using 8 different primer combinations, in total 48 cDNA were differentially expressed between SSc and healthy dermal fibroblasts. Sequence analysis identified distinct PCR products, which were overexpressed in SSc as highly homologous to gene segments of gremlin protein, lysyl oxidase, c-cbl proto-oncogene, an estrogen-responsive element, fibronectin, and collagen type XII alpha 1 precursor. Conclusion. Our results show that RAP-PCR is a suitable method to identify differentially expressed genes in SSc fibroblasts. Further, we identified genes that have not yet been described in the pathophysiology of SSc and that may be involved in matrix synthesis and cellular interaction.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | LYSYL OXIDASE EXPRESSION; COLLAGEN; GREMLIN; SCLERODERMA; FIBROSIS; PROTEIN; SKIN; DNA; AMPLIFICATION; NEPHROPATHY; systemic sclerosis; fibroblasts; gene expression; differential display |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Dermatologie und Venerologie Medicine > Lehrstuhl für Innere Medizin I |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 16 Dec 2020 14:50 |
| Last Modified: | 16 Dec 2020 14:50 |
| URI: | https://pred.uni-regensburg.de/id/eprint/32974 |
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