Differential effects of estrogen receptor beta isoforms on glioblastoma progression

We examined the transcriptional changes modulated by knocking out ERβ and reintroduction of ERβ1 and ERβ5 isoforms by perfroming global transcriptome analysis. ERβ was knocked out in U87 cells using CRISPR/Cas9 system and reintroduced the ERβ1 and ERβ5 isoforms in knockout background. RNA was isolated from control U87, U87-ERβ-KO, U87-ERβ1 and U87-ERβ5 cells and utilized for RNA-seq analysis. Our results demonstrated that ERβ-KO, ERβ1 and ERβ5 modulated unique pathways including NF-κB singaling, Jak/STAT pathway and mTOR signaling. Total RNA was isolated from U87 control, U87-ERβ-KO, U87-ERβ1 and U87-ERβ5 cells. Illumina TruSeq RNA Sample Preparation was performed following manufacturer's protocol. Samples were run on an Illumina HiSeq 3000 in duplicates. The combined raw reads were aligned to UCSC hg19 and genes were annotated by Tophat. Genes were annotated and quantified by HTSeq-DESeq pipeline.