Coordinated transcriptional and catabolic programs support iron dependent adaptation to RAS-MAPK pathway inhibition in pancreatic cancer [RNA-Seq]

In this study, human pancreatic ductal adenocarcinoma cells lines (KP4, MiaPaca2) were treated with the MEK inhibitor, Trametinib or DMSO for 48hrs prior to RNA-seq profiling to evaluate significant gene expression changes associated with MEK inhibition. To assess the role of transcription factors MYC and TFE3 in regulation of gene expression changes associated with MEK inhibition, we performed chromatin immunoprecipitation using antibodies against c-MYC and TFE3 in KP4 cells following 48hrs of Trametinib or DMSO treatment. Overall design: Two human pancreatic cancer cell lines, KP4 and MiaPaca2 were treated with DMSO or Trametinib for 48h in triplicates and the gene expression changes were analyzed by RNA-Sequencing.

本研究中，在进行RNA测序（RNA-seq）分析前，使用MEK抑制剂（MEK inhibitor）曲美替尼（Trametinib）或二甲基亚砜（DMSO）处理人胰腺导管腺癌细胞系（KP4、MiaPaca2）48小时，以评估与MEK抑制相关的显著基因表达变化。为探究转录因子（transcription factors）MYC与TFE3在调控MEK抑制相关基因表达变化中的作用，我们在KP4细胞经曲美替尼或DMSO处理48小时后，使用针对c-MYC和TFE3的抗体进行染色质免疫沉淀（chromatin immunoprecipitation）实验。实验设计概述：选取KP4与MiaPaca2两株人胰腺癌细胞系，分别使用DMSO或曲美替尼处理48小时，每组设置3次生物学重复，随后通过RNA测序（RNA-Sequencing）分析基因表达变化。