Abstract | INTRODUCTION: METHODS: We used real-time reverse transcription-polymerase chain reaction to measure LG268- and rosiglitazone-mediated inhibition of MMP gene transcription in IL-1-beta-treated SW-1353 chondrosarcoma cells. An in vitro collagen destruction assay was a functional readout of MMP collagenolytic activity. Luciferase reporter assays tested the function of a putative regulatory element in the promoters of MMP-1 and MMP-13, and chromatin immunoprecipitation (ChIP) assays detected PPARgamma and changes in histone acetylation at this site. Post-translational modification of RXR and PPARgamma by small ubiquitin-like modifier (SUMO) was assayed with immunoprecipitation and Western blot. RESULTS:
Rosiglitazone inhibited MMP-1 and MMP-13 expression in IL-1-beta-treated SW-1353 cells at the mRNA and heterogeneous nuclear RNA levels and blunted IL-1-beta-induced collagen destruction in vitro. Combining LG268 and rosiglitazone had an additive inhibitory effect on MMP-1 and MMP-13 transcription and collagenolysis. IL-1-beta inhibited luciferase expression in the MMP reporter assay, but rosiglitazone and LG268 had no effect. ChIP indicated that treatment with IL-1-beta, but not LG268 and rosiglitazone, increased PPARgamma at the proximal promoters of both MMPs. Finally, rosiglitazone or LG268 induced 'cross-SUMOylation' of both the target receptor and its binding partner, and IL-1-beta-alone had no effect on SUMOylation of RXR and PPARgamma but antagonized the ligand-induced SUMOylation of both receptors. CONCLUSIONS: The PPARgamma and RXR ligands rosiglitazone and LG268 may act through similar mechanisms, inhibiting MMP-1 and MMP-13 transcription. Combinatorial treatment activates each partner of the RXR: PPARgamma heterodimer and inhibits IL-1-beta-induced expression of MMP-1 and MMP-13 more effectively than either compound alone. We conclude that the efficacy of combined treatment with lower doses of each drug may minimize potential side effects of treatment with these compounds.
|
Authors | Peter S Burrage, Adam C Schmucker, Yanqing Ren, Michael B Sporn, Constance E Brinckerhoff |
Journal | Arthritis research & therapy
(Arthritis Res Ther)
Vol. 10
Issue 6
Pg. R139
( 2008)
ISSN: 1478-6362 [Electronic] England |
PMID | 19046432
(Publication Type: Comparative Study, Journal Article, Research Support, N.I.H., Extramural)
|
Chemical References |
- Drug Combinations
- LG 268
- Matrix Metalloproteinase Inhibitors
- Organic Chemicals
- PPAR gamma
- Retinoid X Receptors
- Thiazolidinediones
- Rosiglitazone
- Matrix Metalloproteinases
|
Topics |
- Animals
- Cattle
- Cell Line, Tumor
- Chondrocytes
(drug effects, enzymology)
- Drug Combinations
- Gene Expression Regulation, Enzymologic
(drug effects, physiology)
- Humans
- Matrix Metalloproteinase Inhibitors
- Matrix Metalloproteinases
(biosynthesis, genetics)
- Organic Chemicals
(administration & dosage)
- PPAR gamma
(agonists, metabolism)
- Retinoid X Receptors
(agonists, metabolism)
- Rosiglitazone
- Thiazolidinediones
(administration & dosage)
|