RNA-seq of shE4F1 or shCTRL SUM159 triple negative breast cancer cell line treated or not with Gemcitabine

E4F1 is a ubiquitously expressed zinc-finger protein of the Gli-Kruppel family that was first identified, more than 30 years ago, as a cellular target of the adenoviral oncoprotein E1A13S (Ad type V), required for transcriptional regulation of adenoviral genes. In this study we investigated the impact of direct translational target genes on DNA damage response induced by Gemcitabine. To do so, we performed RNAseq SUM159 triple negative breast cancer cell line. SUM159 was tranfected either with shE4F1 OR shscramble treated or not with Gemcitabine for 24 hours. mRNA profiles of SUM159 treated or not with Gemcitabine and transfected either with scramble shRNA or shRNA-E4F1 were generated using TruSeq stranded RNA Library Prep Kit (Illumina) followed by sequencing with 50 bp single-end reads on HiSeq 2500.

E4F1是一种广泛表达的Gli-Kruppel家族锌指蛋白（zinc-finger protein），三十余年前首次被鉴定为腺病毒癌蛋白E1A13S（Ad V型）的细胞靶点，该靶点是腺病毒基因转录调控所必需的。本研究旨在探究直接翻译靶基因对吉西他滨（Gemcitabine）诱导的DNA损伤应答的影响。为此，我们对SUM159三阴性乳腺癌细胞系开展了RNA测序（RNAseq）实验：将SUM159细胞分别转染靶向E4F1的短发夹RNA（shE4F1）或阴性对照短发夹RNA（shscramble），并分别用吉西他滨处理或不予处理，孵育24小时。采用TruSeq链特异性RNA文库制备试剂盒（TruSeq stranded RNA Library Prep Kit，Illumina公司）构建测序文库，随后在HiSeq 2500测序平台上以50 bp单端读长进行测序，由此获取经吉西他滨处理/未处理、转染阴性对照shRNA或shRNA-E4F1的SUM159细胞的mRNA表达谱。