The transcriptome effect of knocking down EZH2 in TamR MCF7L

Purpose: Increasing evidence suggests that epigenetic reprogramming contributes significantly to the development of endocrine therapy resistance in breast cancer. The goal of this work is to explore how the histone methyltransferase EZH2 interacts with ER signaling and drives the insensitiveness of breast cancer cells to the antagonistic effect of tamoxifen on ER activity. Therefore, we comprehensively analyzed the transcriptional program regulated by EZH2 in tamoxifen-resistant (TamR) MCF-7 cells. Methods: TamR MCF-7 cells between passage 142-144 upon were used for this assay. For mRNA-Seq, cells are transfected with scrambled control shRNAs (shCtrl) or shRNAs targeting EZH2 (shEZH2). Total RNA were extracted by TRIzol (Invitrogen) and libraries were constructed using Illumina TruSeq RNA Sample Prep Kit v2 (Cat.# RS-122-2001). Hiseq 3000 was used for sequencing. Overall design: Transcriptome profiles of Tamoxifen resistance MCF-7 infected with shCtrl or shEZH2 were sequenced in duplicate using Illumina HiSeq 3000.