Abstract | OBJECTIVE: To examine the effects of specific inhibition of geranylgeranyl transferase I on the expression of types I and III collagen genes in normal and systemic sclerosis (SSc) dermal fibroblasts in vitro. METHODS: Fibroblasts from 2 normal subjects and 4 SSc patients were incubated with 2-10 microM of GGTI-298, a specific geranylgeranyl transferase inhibitor. Type I collagen and fibronectin production were determined by enzyme-linked immunosorbent assay. Steady-state messenger RNA ( mRNA) levels for alpha1(I), alpha2(I), and alpha1(III) collagens and fibronectin were assessed by Northern hybridization, and the transcription of the alpha1(I) collagen gene was examined by transient transfections with a reporter construct containing -5.3 kb of the gene. RESULTS:
GGTI-298 caused a dose-dependent inhibition of type I collagen production and a reduction in the steady-state levels of alpha1(I), alpha2(I), and alpha1(III) mRNA in normal and SSc cells. A 60-70% inhibition of type I collagen production and a 70-80% reduction in the mRNA levels for alpha1(I), alpha2(I), and alpha1(III) were observed at 10 microM GGTI-298. In contrast, the expression of fibronectin, cyclooxygenase 1, and GAPDH was not affected. The effects on alpha1(I) collagen mRNA resulted from a profound reduction in transcription of the alpha1(I) collagen gene promoter. GGTI-298 did not affect cellular viability or morphology. CONCLUSION: These results demonstrate that specific inhibition of geranylgeranyl prenylation causes a potent and selective inhibition of expression of the genes encoding types I and III collagens, without affecting cellular viability. The findings indicate that inhibition of geranylgeranyl prenylation should be further studied as a potential therapeutic approach for SSc and other fibrosing diseases.
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Authors | J Rosenbloom, B Saitta, S Gaidarova, N Sandorfi, J C Rosenbloom, W R Abrams, A D Hamilton, S M Sebti, U Kucich, S A Jimenez |
Journal | Arthritis and rheumatism
(Arthritis Rheum)
Vol. 43
Issue 7
Pg. 1624-32
(Jul 2000)
ISSN: 0004-3591 [Print] United States |
PMID | 10902768
(Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)
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Chemical References |
- Actins
- Benzamides
- Enzyme Inhibitors
- Fibronectins
- GGTI 298
- Isoenzymes
- Membrane Proteins
- RNA, Messenger
- Collagen
- Cyclooxygenase 1
- PTGS1 protein, human
- Prostaglandin-Endoperoxide Synthases
- Alkyl and Aryl Transferases
- geranylgeranyltransferase type-I
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Topics |
- Actins
(drug effects, genetics, metabolism)
- Alkyl and Aryl Transferases
(antagonists & inhibitors)
- Benzamides
(pharmacology)
- Cell Line
- Cell Line, Transformed
- Collagen
(drug effects, genetics, metabolism)
- Cyclooxygenase 1
- Enzyme Inhibitors
(pharmacology)
- Fibroblasts
(enzymology, pathology)
- Fibronectins
(metabolism)
- Humans
- Isoenzymes
(metabolism)
- Membrane Proteins
- Prostaglandin-Endoperoxide Synthases
(metabolism)
- RNA, Messenger
(metabolism)
- Scleroderma, Systemic
(enzymology, pathology)
- Skin
(enzymology, pathology)
- Transcription, Genetic
(drug effects)
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