Lengthy noncoding RNAs (lncRNA) have been associated with the development of cancer

Lengthy noncoding RNAs (lncRNA) have been associated with the development of cancer. a regulator of apoptosis (21). was characterized as a novel prostate-specific lncRNA, regulator of cell proliferation, and target of the polycomb repressive complex 2 (PRC2) (15). RNA binds to AR protein to block the interaction with the E3 ubiquitin ligase MDM2, thereby preventing protein degradation and AR activation (22). In a previous study, we analyzed global AR transcriptional network by mapping genome-wide transcriptional start 6-TAMRA sites regulated by androgen and AR binding sites (ARBS). This integrative genomic study revealed comprehensive AR-regulated transcripts from intergenic or AS regions of genes in prostate cancer cells (23). In addition, we investigated the functional roles of these novel androgen-responsive long noncoding RNAs, such as a lncRNA located at the AS region of the C-terminal-binding protein 1 promoted cell growth and migration and repressed several genes related to the apoptosis pathway, including would play an important role in the progression of prostate cancer. Results Identification of Androgen-induced lncRNAs by Directional RNA Sequencing To investigate hormone-regulated lncRNAs in prostate cancer, we performed directional RNA sequencing (RNA-seq) and identified lncRNAs induced by androgen in prostate cancer cell lines. For lncRNA analysis, we used two databases, GENCODE V19 (25) and NONCODE v4. AR-positive prostate cancer cell lines, LNCaP and VCaP, and their corresponding castration-resistant cell lines, LTAD and VCaP-LTAD (24), were treated with vehicle (ethanol) or 10 nm 5-dihydrotestosterone (DHT). In addition, LNCaP and VCaP cells were also treated with DHT plus bicalutamide or with 10 nm siRNA-targeting AR (siAR). After 24 h, total RNAs were extracted, and then RNA-seq analysis was performed. Bioinformatic analysis identified lncRNAs that were up-regulated more than 1.5-fold by DHT treatment and repressed to less than 0.75-fold by bicalutamide and siAR treatment in both LNCaP and VCaP cell lines. Nine transcripts were common in both cell lines using the GENCODE annotation and two in the NONCODE annotation (Fig. 1as a lncRNA highly expressed in castration-resistant prostate cancer. as an AR-targeted lncRNA up-regulated in LTAD cells. Venn diagram representing overlap of lncRNAs induced by androgen (10 nm DHT) and repressed by 6-TAMRA 1 mm bicalutamide and 10 nm siAR treatment for 24 h in LNCaP and VCaP cell lines. = 3. Expression levels are presented relative to the value of as the reference gene. Values represent mean S.D. TABLE 1 List of androgen-induced lncRNAs lncRNAs induced by more than 1.5-fold with 10 nm DHT compared with ethanol (Et) treatment and repressed to less than 0.75-fold by bicalutamide (Bic) and siAR in LNCaP and VCaP cell lines are listed. RPKM, reads per kilobase per million mapped reads. Open in a separate window SOCS2-AS1 Is an Androgen-induced lncRNA Highly Expressed in Castration-resistant Prostate Cancer Cells Next, we performed qRT-PCR to analyze the 6-TAMRA expression of five lncRNAs in both LNCaP and VCaP and their LTAD cells. We validated their androgen induction as observed in RNA-seq data (Fig. 1, and was highly expressed in LTAD and VCaP-LTAD compared with the parental cell lines by RNA-seq and qRT-PCR analysis (Figs. 1, and and in LNCaP and VCaP cell lines treated with 10 nm DHT or ethanol (and mRNA after DHT treatment in LNCaP and LTAD cell lines determined by qRT-PCR. 0.01. is an antisense lncRNA transcribed from the opposite strand of the protein coding the gene. is one of the eight members of Rabbit Polyclonal to Retinoic Acid Receptor alpha (phospho-Ser77) the SOCS family that are induced by cytokine stimulation through the Janus kinase (JAK/STAT) signaling. genes contribute to cytokine inhibition by reducing JAK or STAT phosphorylation, inhibiting the same cascade that initiated.