Towards resolution of the intron retention paradox in breast cancer

We explored intron retention patterns in normal breast epithelial cells (MCF10A) and estrogen receptor positive (ER+) breast cancer cells (MCF7). Total RNA was isolated from MCF7 and MCF10A cells using Trizol (Invitrogen). The RNA quality was assessed using RNA 6000 Nano Chips on an Agilent Bioanalyzer (Agilent Technologies) to confirm an RNA integrity score of >7.0. mRNA-seq was performed by Macrogen (Korea) using the Illumina Hi-Seq 2000 platform. RNA-seq libraries were prepared from > 1 μg of total RNA using TruSeq RNA sample prep kit (Illumina) according to the manufacturers’ instructions.

本研究针对正常乳腺上皮细胞（MCF10A）与雌激素受体阳性（ER+）乳腺癌细胞（MCF7）的内含子保留（intron retention）模式展开探究。从MCF7及MCF10A细胞中采用Trizol试剂（Invitrogen公司）提取总RNA；通过安捷伦生物分析仪（Agilent Technologies）搭配RNA 6000 Nano芯片对RNA质量进行检测，确认RNA完整性评分大于7.0。mRNA测序（mRNA-seq）由韩国Macrogen公司基于Illumina Hi-Seq 2000测序平台完成；取≥1 μg总RNA，严格遵循试剂盒说明书，使用TruSeq RNA样本制备试剂盒（Illumina公司）完成RNA-seq文库构建。