Androgen receptor (AR) expression in prostate cancer (CaP) cells varies due to the multiple changes including epigenetic modifications such as DNA methylation and histone deacetylation. DNA methyltransferase and histone deacetylase inhibitors are promising for the treatment of CaP. The aim of our study was to analyze the 5-aza-2'-deoxycytidine (Aza‑dC) and sodium butyrate (NaB) effects on CaP cells with modified AR gene expression. The androgen-independent human prostate cancer cell lines PC3 (lacking a functional AR) and DU145 (strongly limited expression due to methylations in the AR gene) were used. PCR of bisulfite-modified DNA and RT-PCR with bisulfite-sequencing were used for AR gene analysis of DU145 and PC3 cells following their treatment with Aza-dC and/or NaB. Re-acetylated histones around the AR gene were detected by conventional PCR of immunoprecipitated DNA obtained from treated cells. In both cell lines without the AR expression, the combined treatment was followed with significant decrease of cell viability. The co-treatment of DU145 cells caused site-specific demethylation in the AR promoter region followed by gene re-expression and increased acetylation in histones H3 and H4. The co-treatment with Aza-dC and NaB was the most effective in demethylation and re-expression of the AR gene. In the AR gene promoter, the location and density of deme-thylated CpGs indicated the existence of distinct promoter hot spot that could be a target of AR gene inactivation therapy of CaP patients during androgen deprivation.

• MeSH
• 5' nepřekládaná oblast MeSH
• acetylace účinky léků   MeSH
• androgenní receptory biosyntéza   genetika   metabolismus   MeSH
• androgeny genetika   metabolismus   MeSH
• azacytidin analogy a deriváty   farmakologie   MeSH
• DNA genetika   MeSH
• histondeacetylasy genetika   metabolismus   MeSH
• histony genetika   metabolismus   MeSH
• inhibitory histondeacetylas farmakologie   MeSH
• kyselina máselná farmakologie   MeSH
• lidé MeSH
• methyltransferasy antagonisté a inhibitory   genetika   metabolismus   MeSH
• metylace DNA účinky léků   MeSH
• nádorové buněčné linie MeSH
• nádory prostaty rezistentní na kastraci farmakoterapie   enzymologie   genetika   metabolismus   MeSH
• promotorové oblasti (genetika) účinky léků   genetika   MeSH
• viabilita buněk účinky léků   genetika   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Sodium butyrate, as a naturally occurring inhibitor of histone deacetylases (HDACI), is a non-toxic agent, with an ability to change histone acetylation and expression of large number genes. This study shows different effects of sodium butyrate on expression and transcription activity of the androgen receptor in cancer (LNCaP, C4-2) and normal (RWPE-1) prostate cells. Moreover, we studied the coregulator expressions and histone acetylation alteration in cancer and normal cells. Coregulators, coactivators as well as corepressors, play an important role in AR-mediated growth and progression of prostate cancer. There is a competition between coactivators and corepressors for binding on the AR and therefore the changes in coregulators expression and ratio could be important for prostate cancer survival. Our study was focused on two coregulators, SMRT and p300, which interact with AR in multiprotein complex and affect the AR transcription activity. Our data indicate that sodium butyrate has an effect on AR coregulators expression, transcription activity and histone acetylation in cancer cells, but there is only minimal effect in normal cells. In addition, the results of changes in acetylation level on lysine residues of histone H4 after sodium butyrate treatment confirm its epigenetic effect on prostate cancer cells.

• MeSH
• acetylace MeSH
• androgenní receptory genetika   metabolismus   MeSH
• buněčné linie MeSH
• histony metabolismus   MeSH
• inhibitory histondeacetylas farmakologie   MeSH
• kyselina máselná farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory prostaty metabolismus   MeSH
• prostata cytologie   MeSH
• prostatický specifický antigen metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• androgenní receptory účinky léků   MeSH
• buněčné linie cytologie   účinky léků   MeSH
• butyráty farmakologie   MeSH
• financování organizované MeSH
• nádorové buněčné linie cytologie   účinky léků   MeSH
• nádory prostaty farmakoterapie   MeSH
• techniky in vitro MeSH
• viabilita buněk účinky léků   MeSH

• MeSH
• butyráty aplikace a dávkování   farmakologie   MeSH
• dospělí MeSH
• financování organizované MeSH
• lidé MeSH
• nádorové buněčné linie účinky léků   MeSH
• techniky in vitro MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH

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. Furthermore, we estimated the reduced presence of HDAC2 and HDAC3 proteins by NaB and TSA treatment in AR-negative DU145 cell line. 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 protein.

• MeSH
• androgenní receptory genetika   metabolismus   MeSH
• butyráty metabolismus   farmakologie   terapeutické užití   MeSH
• časové faktory MeSH
• histondeacetylasa 2 metabolismus   MeSH
• histondeacetylasy metabolismus   MeSH
• imunoprecipitace MeSH
• inhibitory histondeacetylas terapeutické užití   MeSH
• korepresor 2 jaderného receptoru genetika   metabolismus   MeSH
• kyseliny hydroxamové metabolismus   farmakologie   terapeutické užití   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory prostaty farmakoterapie   genetika   metabolismus   MeSH
• regulace genové exprese u nádorů MeSH
• vazba proteinů genetika   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH