Skip to main content
. 2014 Nov 21;5:5383. doi: 10.1038/ncomms6383

Figure 2. ERα regulated NEAT1 lncRNA is upregulated in prostate cancer.

Figure 2

(a) NEAT1 is overexpressed in various prostate data sets (Oncomine). (b) Distribution of the median expression of all genes (core transcript clusters) on the Human Exon 1.0 ST array in the pooled Mayo Clinic cohort (n=594). NEAT1’s expression ranks in the 99th percentile of all genes on the array. (c) Expression of NEAT1 with/without ERα overexpression and E2 treatment (10 nM) at different time points in a panel of prostate cancer cell lines. Results are expressed as the mean±s.d. of three independent experiments. (d) View of NEAT1 genomic location indicates presence of two ERα-binding sites in the promoter region. Read coverage tracks derived from RNA-sequencing data indicates a higher abundance of NEAT1 transcripts in PCa compared with benign tumours in three representative cases. The figure also reports the ChIP-sequencing coverage tracks for ERα (VCaP ERα, VCaP and input DNA as control). The bottom panel shows the binding sites of ERα, AR (GEO Accession GSM353651-tissue AR (ref. 25)), Ace-H3, H3K4me3 and H3K36me3 in VCaP cell line (GEO Accession GSM353629, GSM353620 and GSM353624 (ref. 25)), respectively. (e) Chromatin immunoprecipitation followed by qPCR to study ERα recruitment to NEAT1 promoter in VCaP cells with/without E2 treatment (10 nM) was performed with primers spanning the binding regions identified by ERα ChIP-seq data. Primers for nonspecific region were used as negative control for ChIP studies. Results are expressed as percentage of input from two independent experiments. Vertical error bars represent the range of data. (f) Luciferase-based promoter reporter assays was used to analyse effect of ERα and/or AR on ERE-Luc promoter in VCaP cells. Cells were transiently transfected with the (ERE)3-SV40-luc reporter plasmid and ERα, or AR-treated with/without E2 or R1881 (1 nM) for 48 h. Results are expressed as the mean±s.d calculated from three independent experiments. (g) Luciferase-based promoter reporter assays were used to analyse NEAT1 promoter activity following ERα expression −/+E2 (10 nM) for 24 h. Results are expressed as the means±s.d. calculated from three independent experiments. Student’s t-test was performed for comparisons where indicated, and *P<0.05 and **P<0.01 were considered statistically significant.