Transcriptome profiling of MDA-MB-231 cells following the MEL-18 phosphorylation at T334

RNA-sequencing analysis of MEL-18 WT- or MEL-18 T334A-overexpressing MDA-MB-231 cell lines. MEL-18, a core component of polycomb-repressive complex (PRC)-1, has been known to be phosphorylated at multiple residues in vitro; however, its functional roles in mammalian cells and human cancer remains largely unknown. Here, we examined the effect of MEL-18 phosphorylation at T334 site on polycomb-mediated epigenetic silencing in human breast cancer. Our results demonstrated that the phosphorylation of MEL-18 at T334 alters its genomic distribution and transcriptional activity that reflects functional change of MEL-18 in modulating breast tumour progression. MDA-MB-231 cells stably expressing either MEL-18 wild type (WT) or T334 phosphorylation-deficient mutant form of MEL-18 (MEL-18 T334A) and control cells (CON, empty vector) were cultured in DMEM supplemented with 10% FBS for 48h. Total RNA was isolated from the cultures using Trizol reagent. For each of the 3 conditions, 3 biological replicates were included. In total, 9 RNA-sequencing samples were analyzed; 3 controls, 3 MEL-18 WT, and 3 MEL-18 T334A overexpressing cells.

本数据集针对过表达MEL-18野生型（WT）或MEL-18 T334A突变体的MDA-MB-231细胞系开展RNA测序（RNA-sequencing）分析。MEL-18是多梳抑制复合体1（polycomb-repressive complex 1, PRC1）的核心组分，此前已知其在体外可被多个位点磷酸化；然而其在哺乳动物细胞及人类癌症中的功能作用仍在很大程度上未被阐明。本研究探究了T334位点的MEL-18磷酸化对人类乳腺癌中多梳复合体介导的表观遗传沉默的影响，结果显示MEL-18在T334位点的磷酸化会改变其基因组分布与转录活性，这反映了MEL-18在调控乳腺肿瘤进展过程中的功能变化。实验中将稳定表达MEL-18野生型（WT）、T334磷酸化缺陷型MEL-18突变体（MEL-18 T334A）的MDA-MB-231细胞，以及对照细胞（CON，空载体）置于添加了10%胎牛血清（fetal bovine serum, FBS）的达尔伯克改良伊格尔培养基（Dulbecco's Modified Eagle Medium, DMEM）中培养48小时，使用Trizol试剂从培养物中提取总RNA。每组实验条件均设置3次生物学重复，共分析9个RNA测序样本，包括3个对照样本、3个过表达MEL-18野生型的样本以及3个过表达MEL-18 T334A的样本。