Cargando…
PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling
BACKGROUND: PCA3 is a non-coding RNA (ncRNA) that is highly expressed in prostate cancer (PCa) cells, but its functional role is unknown. To investigate its putative function in PCa biology, we used gene expression knockdown by small interference RNA, and also analyzed its involvement in androgen re...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2012
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3544699/ https://www.ncbi.nlm.nih.gov/pubmed/23130941 http://dx.doi.org/10.1186/1471-2407-12-507 |
_version_ | 1782255831773872128 |
---|---|
author | Ferreira, Luciana Bueno Palumbo, Antonio de Mello, Kivvi Duarte Sternberg, Cinthya Caetano, Mauricio S de Oliveira, Felipe Leite Neves, Adriana Freitas Nasciutti, Luiz Eurico Goulart, Luiz Ricardo Gimba, Etel Rodrigues Pereira |
author_facet | Ferreira, Luciana Bueno Palumbo, Antonio de Mello, Kivvi Duarte Sternberg, Cinthya Caetano, Mauricio S de Oliveira, Felipe Leite Neves, Adriana Freitas Nasciutti, Luiz Eurico Goulart, Luiz Ricardo Gimba, Etel Rodrigues Pereira |
author_sort | Ferreira, Luciana Bueno |
collection | PubMed |
description | BACKGROUND: PCA3 is a non-coding RNA (ncRNA) that is highly expressed in prostate cancer (PCa) cells, but its functional role is unknown. To investigate its putative function in PCa biology, we used gene expression knockdown by small interference RNA, and also analyzed its involvement in androgen receptor (AR) signaling. METHODS: LNCaP and PC3 cells were used as in vitro models for these functional assays, and three different siRNA sequences were specifically designed to target PCA3 exon 4. Transfected cells were analyzed by real-time qRT-PCR and cell growth, viability, and apoptosis assays. Associations between PCA3 and the androgen-receptor (AR) signaling pathway were investigated by treating LNCaP cells with 100 nM dihydrotestosterone (DHT) and with its antagonist (flutamide), and analyzing the expression of some AR-modulated genes (TMPRSS2, NDRG1, GREB1, PSA, AR, FGF8, CdK1, CdK2 and PMEPA1). PCA3 expression levels were investigated in different cell compartments by using differential centrifugation and qRT-PCR. RESULTS: LNCaP siPCA3-transfected cells significantly inhibited cell growth and viability, and increased the proportion of cells in the sub G0/G1 phase of the cell cycle and the percentage of pyknotic nuclei, compared to those transfected with scramble siRNA (siSCr)-transfected cells. DHT-treated LNCaP cells induced a significant upregulation of PCA3 expression, which was reversed by flutamide. In siPCA3/LNCaP-transfected cells, the expression of AR target genes was downregulated compared to siSCr-transfected cells. The siPCA3 transfection also counteracted DHT stimulatory effects on the AR signaling cascade, significantly downregulating expression of the AR target gene. Analysis of PCA3 expression in different cell compartments provided evidence that the main functional roles of PCA3 occur in the nuclei and microsomal cell fractions. CONCLUSIONS: Our findings suggest that the ncRNA PCA3 is involved in the control of PCa cell survival, in part through modulating AR signaling, which may raise new possibilities of using PCA3 knockdown as an additional therapeutic strategy for PCa control. |
format | Online Article Text |
id | pubmed-3544699 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-35446992013-01-16 PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling Ferreira, Luciana Bueno Palumbo, Antonio de Mello, Kivvi Duarte Sternberg, Cinthya Caetano, Mauricio S de Oliveira, Felipe Leite Neves, Adriana Freitas Nasciutti, Luiz Eurico Goulart, Luiz Ricardo Gimba, Etel Rodrigues Pereira BMC Cancer Research Article BACKGROUND: PCA3 is a non-coding RNA (ncRNA) that is highly expressed in prostate cancer (PCa) cells, but its functional role is unknown. To investigate its putative function in PCa biology, we used gene expression knockdown by small interference RNA, and also analyzed its involvement in androgen receptor (AR) signaling. METHODS: LNCaP and PC3 cells were used as in vitro models for these functional assays, and three different siRNA sequences were specifically designed to target PCA3 exon 4. Transfected cells were analyzed by real-time qRT-PCR and cell growth, viability, and apoptosis assays. Associations between PCA3 and the androgen-receptor (AR) signaling pathway were investigated by treating LNCaP cells with 100 nM dihydrotestosterone (DHT) and with its antagonist (flutamide), and analyzing the expression of some AR-modulated genes (TMPRSS2, NDRG1, GREB1, PSA, AR, FGF8, CdK1, CdK2 and PMEPA1). PCA3 expression levels were investigated in different cell compartments by using differential centrifugation and qRT-PCR. RESULTS: LNCaP siPCA3-transfected cells significantly inhibited cell growth and viability, and increased the proportion of cells in the sub G0/G1 phase of the cell cycle and the percentage of pyknotic nuclei, compared to those transfected with scramble siRNA (siSCr)-transfected cells. DHT-treated LNCaP cells induced a significant upregulation of PCA3 expression, which was reversed by flutamide. In siPCA3/LNCaP-transfected cells, the expression of AR target genes was downregulated compared to siSCr-transfected cells. The siPCA3 transfection also counteracted DHT stimulatory effects on the AR signaling cascade, significantly downregulating expression of the AR target gene. Analysis of PCA3 expression in different cell compartments provided evidence that the main functional roles of PCA3 occur in the nuclei and microsomal cell fractions. CONCLUSIONS: Our findings suggest that the ncRNA PCA3 is involved in the control of PCa cell survival, in part through modulating AR signaling, which may raise new possibilities of using PCA3 knockdown as an additional therapeutic strategy for PCa control. BioMed Central 2012-11-06 /pmc/articles/PMC3544699/ /pubmed/23130941 http://dx.doi.org/10.1186/1471-2407-12-507 Text en Copyright ©2012 Ferreira et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Ferreira, Luciana Bueno Palumbo, Antonio de Mello, Kivvi Duarte Sternberg, Cinthya Caetano, Mauricio S de Oliveira, Felipe Leite Neves, Adriana Freitas Nasciutti, Luiz Eurico Goulart, Luiz Ricardo Gimba, Etel Rodrigues Pereira PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling |
title | PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling |
title_full | PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling |
title_fullStr | PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling |
title_full_unstemmed | PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling |
title_short | PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling |
title_sort | pca3 noncoding rna is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3544699/ https://www.ncbi.nlm.nih.gov/pubmed/23130941 http://dx.doi.org/10.1186/1471-2407-12-507 |
work_keys_str_mv | AT ferreiralucianabueno pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling AT palumboantonio pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling AT demellokivviduarte pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling AT sternbergcinthya pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling AT caetanomauricios pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling AT deoliveirafelipeleite pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling AT nevesadrianafreitas pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling AT nasciuttiluizeurico pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling AT goulartluizricardo pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling AT gimbaetelrodriguespereira pca3noncodingrnaisinvolvedinthecontrolofprostatecancercellsurvivalandmodulatesandrogenreceptorsignaling |