Preclinical Evaluation of Dual mTOR inhibitor, AZD2014, and Castration in Prostate Cancer

ABSTRACT: Purpose: RNA sequence analysis identified active proliferation of T cells in the combination therapy Methods: Pten-null tumours were collected 4 hours after the last dose (18th day). Total RNA from tumor samples was isolated using the Allprep kit (Qiagen, Germany). The quantity of the RNA was assessed by Qubit (Life technologies) and quality was measured by Bioanalyzer 2100 (Agilent). Samples with 8 or more RNA integrity number (RIN) were used for library preparation and subsequent RNA sequencing. Library preparation was carried out using the Illumina® TruSeq Stranded mRNA Library Preparation Kit (Life Technologies) according to manufacturer recommendations. Single-end 50 bp sequencing reads were generated using Illumina HiSeq and were aligned to the human genome version hg19 or the mouse genome version mm10 using TopHat v2.0.4. Normalisation of read counts and identification of differential expression of genes (DEGs) were carried out using the DESeq v1.10.1 work flow. Results: RNA sequence analysis identified differential gene expression between treatment groups and combination therapy clearly demonstrated active proliferation of cytotoxic lymphocytes. Conclusions: Our study represents the first detailed analysis of Pten-null transcriptomes, with biologic replicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. Our results show that NGS offers a comprehensive and more accurate quantitative and qualitative evaluation of mRNA content within a cell or tissue. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions.

ORGANISM(S): Mus musculus

PROVIDER: GSE93897 | GEO | 2020/01/21

REPOSITORIES: GEO