Analysis of gene expression in Tgif mutant mouse colon tumors.. Analysis of gene expression in Tgif mutant mouse colon tumors.

Colon tumors in mice were generated by deleting one allele of the Apc gene, or both alleles of Tgif1 and Tgif2, together with a single Apc allele. Gene expression was compared by RNA-seq analysis of wild type colon, Apc mutant tumors or Apc mutant colon tumors with deletion of both Tgif1 and Tgif2. Comparison of Apc tumors to Apc;Tgif1;Tgif2 tumors was used to identify Tgif-dependent gene expression changes associated with intestinal tumorigenesis. Overall design: Tissues were isolated from 12 week old mice. Three groups were analyzed: 1. Wild type colon tissue from three female and two male mice. 2. Colon tumors from Apc mutant mice (generated using Villin-Cre to delete one allele of the Apc gene), with individual tumors isolated from three female and four male mice. 3. Colon tumors from Apc mutant mice with deletion of both Tgif1 and Tgif2 (generated using Villin-Cre to delete one allele of the Apc gene and both alleles each of Tgif1 and Tgif2), with single tumors isolated from three female and four male mice. RNA-seq was used to compare gene expression profiles between wild type colon and the two different geneotype colon tumors.

本研究通过两种方式构建小鼠结肠肿瘤模型：其一为敲除Apc基因（Apc）的一个等位基因，其二为同时敲除Tgif1基因（Tgif1）与Tgif2基因（Tgif2）的两个等位基因，并保留一个Apc等位基因。采用RNA测序（RNA-seq）分析野生型结肠组织、Apc突变型结肠肿瘤，以及同时敲除Tgif1和Tgif2双等位基因的Apc突变型结肠肿瘤的基因表达谱以开展组间比较。通过对比Apc突变型肿瘤与双敲除Tgif1、Tgif2的Apc突变型肿瘤的基因表达差异，筛选出与肠道肿瘤发生相关的、受Tgif调控的基因表达变化。

实验设计：组织样本取自12周龄小鼠，共分为三组：
1. 野生型结肠组织：供体为3只雌性与2只雄性小鼠；
2. Apc突变型结肠肿瘤：通过Villin-Cre系统敲除一个Apc等位基因构建模型，单个肿瘤样本分别取自3只雌性与4只雄性小鼠；
3. 双敲除Tgif1、Tgif2的Apc突变型结肠肿瘤：通过Villin-Cre系统敲除一个Apc等位基因，并分别敲除Tgif1与Tgif2的两个等位基因构建模型，单个肿瘤样本分别取自3只雌性与4只雄性小鼠。
本研究采用RNA-seq比较野生型结肠组织与两种不同基因型结肠肿瘤的基因表达谱。