Signaling through the androgen receptor (AR) plays a critical role in prostate cancer progression. The AR is a classical nuclear receptor (NR) providing a link between signaling molecule and transcription response. Histone deacetylase inhibitors- (HDACI) have antiproliferative and proapoptotic effects on prostate cancer cells and their implication in silence AR signaling may have potential therapeutic use. We aimed to study the inhibitory effects of the corepressor SMRT (Silencing Mediator for Retinoid and Thyroid -hormone receptors) which forms a complex together with nuclear receptor corepressor (N-CoR) and with histone deacetylase 3 (HDAC3) on AR activity.The androgen-sensitive prostate cancer cell line LNCaP and androgen-insensitive prostate cancer cell line C4-2 both AR-positive, and androgen-insensitive DU145 and PC3 prostate cancer cell lines were treated with two HDACIs, sodium butyrate (NaB) and/or trichostatin A (TSA). We amplified immunoprecipitated DNA by conventional PCR and in the -following step we used the chromatin immunoprecipitation (ChIP) analysis coupled with quantitative PCR for monitoring NaB induced formation of AR-SMRT/N-CoR complex binding on the PSA promoter. The co-immunoprecipitation assay revealed increase in AR-SMRT formation in NaB treated cells. Simultaneously, the Western blot analysis showed a significant decrease in AR protein expression. In conclusion, the inhibitory effect of NaB on AR gene expression seems to be specific and unique for prostate cancer AR-positive cell lines and corresponds with its ability to stimulate AR-SMRT complex formation. We suggest that AR and SMRT/N-CoR corepressors may form a stable complex in vitro and NaB may facilitate the interaction between AR nuclear steroid receptor and SMRT corepressor prote.
- MeSH
- androgenní receptory metabolismus MeSH
- butyráty farmakologie MeSH
- histondeacetylasa 2 analýza MeSH
- histondeacetylasy analýza metabolismus MeSH
- inhibitory histondeacetylas terapeutické užití MeSH
- korepresor 1 jaderného receptoru metabolismus MeSH
- kyseliny hydroxamové farmakologie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory prostaty farmakoterapie metabolismus MeSH
- promotorové oblasti (genetika) MeSH
- prostatický specifický antigen genetika MeSH
- proteiny spojené s mnohočetnou rezistencí k lékům metabolismus MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
Přeruš. str. : il., tab. ; 30 cm
Using DNA microarrays, gene expression will be studied in leukemia, in control cell populations and in cell cultures. Changes will be correlated to epigenetic characteristics of chromatin at the level of its modifications (methylation, acetylation). Structural characteristics of chromatin at sites with altered expression will be studied using fluorescence in situ hybridization (FISH), spectral microscopy and immunoFISH. Drugs with narrow spectrum of genes with altered expression wil be studied. Using advanced techniques of gene or protein visualization and top level confocal microscopy, the mechanisms of the induction and progression of leukemia will be investigated with the goal to suggest appropriate markers for diagnostics. We hope that our study ofgene expression in parallel to epigenetics will allow us to propose target molecules of effective therapy.
Bude studována exprese pomocí DNA microarrays u leukémií, vhodných kontrolních buněk a pro in vitro populace. Změny budou korelovány s epigenetickými charakteristikami chromatinu na úrovni jeho modifikací (metylace, acetylace histonů, metylace DNA). Strukturální charakteristiky chromatinu budou zkoumány v místech se změnami exprese pomocí fluorescenční in situ hybridizace, spektrální mikroskopie a imunoFISH. Budou testovány protinádorové látky s úzkým spektrem účinku na úrovni exprese. Využitím pokročilých metod vizualizace jaderných stuktur a moderní konfokální mikroskopie bude studován mechanismus vzniku a progrese leukémií. Cílem bude navrhnout vhodné markry pro diagnostiku. Domníváme se, že studium genů měnících expresi a epigenetických mechanismůstojících za těmito změnami nám umožní navrhnout cílové molekuly pro léčbu.
