Gene expression of SLNs (Synthetic Lethal with Nutlin-3) on HCT116 cells. Homo sapiens

A subset of our SLNs would be upregulated by Nutlin-3 and down-regulated by 5-FU and that this differential regulation could potentially explain how cell fate choice is determined SLNs as a group were largely members of pathways parallel to p53, in the sense that very few of them displayed significant changes in mRNA expression after Nutlin-3 or 5FU treatment Overall design: RNA was harvested from cells treated with Nutlin-3, 5FU, or DMSO, for 12 hours using the RNeasy kit (Qiagen). cDNAs were synthesized and labeled with the Genechip Whole Transcript Sense Target Labeling Kit (Affymetrix) per the manufacturer’s instructions and hybridized to Human Exon 1.0 ST arrays (AffyMetrix)

我们的SLNs亚群可被Nutlin-3上调，同时被5-氟尿嘧啶（5-FU）下调，且这种差异调控或可解释细胞命运选择的决定机制。作为一个整体，SLNs大多属于与p53平行的通路成员，具体表现为：经Nutlin-3或5-FU处理后，仅有极少SLNs的信使核糖核酸（mRNA）表达量出现显著变化。实验设计概述：采用RNeasy试剂盒（Qiagen）提取经Nutlin-3、5-FU或二甲基亚砜（DMSO）处理12小时的细胞总RNA。按照制造商的操作说明，使用Genechip全转录正义靶标标记试剂盒（Affymetrix）合成互补DNA（cDNA）并完成标记，随后将标记产物杂交至人类外显子1.0 ST芯片（AffyMetrix）