Abstract

Prostate cancer (PCa) is the most frequent cancer in North American men and PCa cells rely on the androgen receptor (AR) for growth and survival. To understand the effect of AR in cancer cells, we have treated LNCaP and LAPC4 cells, two immortalized human PCa cells in vitro, with the synthetic androgen R1881 and then performed RNA-seq analyses. High quality sequencing data have been analyzed using our bioinformatic pipeline which consists of FastQC for quality controls, Trimmomatic for trimming, and Kallisto for pseudoalignment to the transcriptome. Differentially expressed genes were identified using DESeq2 after adjustment for false-discovery rate (FDR q values < 0.05) and Relative Log Expression (RLE) normalization. Gene Set Enrichment Analysis (GSEA) was also performed to identify biological pathways significantly modulated by androgens. GSEA analyses identified the androgen signaling pathway, as well as several metabolic pathways, as significantly enriched following androgen stimulation. These analyses highlight the most significant metabolic pathways up-regulated following AR activation. Raw and processed RNA-seq data were deposited and made publicly available on the Gene Expression Omnibus (GEO; GSE128749). These data have been incorporated in a recent article describing the functions of AR as a master regulator of PCa cell metabolism. For more details about interpretation of these results, please refer to “Functional genomics studies reveal the androgen receptor as a master regulator of cellular energy metabolism in prostate cancer” by Gonthier et al. (doi: 10.1016/j.jsbmb.2019.04.016).

Highlights

  • Raghavendra Tejo Karthik Poluri a, Charles Joly Beauparlant a, Arnaud Droit a, Etienne Audet-Walsh a, b, *

  • Prostate cancer (PCa) is the most frequent cancer in North American men and PCa cells rely on the androgen receptor (AR) for growth and survival

  • Gene Set Enrichment Analysis (GSEA) was performed to identify biological pathways significantly modulated by androgens

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Summary

Sequencing

Excellent RNA integrity was confirmed using a TapeStation 2200 (Agilent); all samples had an RNA integrity number equivalent (RINe) > 8.5. mRNA enrichment and library preparation were performed using the NEBNext Ultra II Directional RNA library prep kit following the manufacturer's protocol. RNA was sent to the Genomic Centre of the Centre de recherche du CHU de Quebec - Universite Laval for sequencing using a HiSeq 2500 (125bp paired-end sequencing)

RNA-seq analysis
Differential gene expression and GSEA analysis
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