PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling.
PCA3 noncoding RNA is involved in the control of prostate-cancer cell survival and modulates androgen receptor signaling.
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DOI：
10.1186/1471-2407-12-507
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发表时间：
2012-11-06
期刊：
影响因子：
3.8
通讯作者：
Gimba ER
中科院分区：
文献类型：
--
作者：
Ferreira LB;Palumbo A;de Mello KD;Sternberg C;Caetano MS;de Oliveira FL;Neves AF;Nasciutti LE;Goulart LR;Gimba ER
PCA3是一种非编码RNA(NcRNA)，在前列腺癌(Pca)细胞中高表达，但其功能尚不清楚。为了研究它在前列腺癌生物学中的可能功能，我们利用小干扰RNA抑制基因表达，并分析了它在雄激素受体(AR)信号转导中的作用。以LNCaP和PC3细胞为体外模型，针对PCA3外显子4设计了3种不同的siRNA序列。实时荧光定量聚合酶链式反应(Real-time qRT-PCR)和细胞生长、存活和凋亡检测分析了转基因细胞。用100 nM双氢睾酮(DHT)及其拮抗剂(氟他胺)处理LNCaP细胞，并分析一些AR调节基因(TMPRSS2、NDRG1、GREB1、PSA、AR、FGF8、CDK1、CDK2和PMEPA1)的表达，研究PCA3与雄激素受体(AR)信号通路的关系。采用差速离心法和定量逆转录聚合酶链式反应(qRT-PCR)检测PCA3在不同细胞室的表达水平。与干扰siRNA(SiSCr)组相比，LNCaP siPCA3组显著抑制细胞生长和活力，增加细胞周期中处于亚G0/G1期的细胞比例和固缩细胞核百分比。DHT处理的LNCaP细胞PCA3表达显著上调，该作用可被氟他胺逆转。在siPCA3/LNCaP转染组中，AR靶基因的表达较siSCr转染组下调。SiPCA3基因的导入也抵消了DHT对AR信号级联反应的刺激作用，显著下调了AR靶基因的表达。对不同细胞室中PCA3表达的分析表明，PCA3的主要功能存在于细胞核和微体细胞组分中。我们的发现表明，ncRNA PCA3参与了对PCa细胞存活的控制，部分是通过调节AR信号，这可能为将PCA3基因敲除作为一种额外的治疗策略用于PCa控制提供新的可能性。
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. 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. 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. 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.
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