Gene expression profiling of Smad2/3 cKO mice. Gene expression profiling of Smad2/3 cKO mice

Uterine double conditional inactivation of Smad2 and Smad3 in mice results in endometrial dysregulation, infertility, and uterine cancer. Smad2/3 cKO mice demonstrate abnormal expression of genes involved in inflammation, cell-cycle checkpoint, migration, steroid biosynthesis, and SMAD1/5-driven genes. We performed RNA-sequencing to identify the gene expression differences between the uterine epithelium of control and Smad2/3 cKO. To control for estrous cycle variations, the uterine epithelium was collected from mice at 0.5 dpc. Global gene expression profiles of Smad2/3 cKO versus control mice was analyzed. Our RNA sequencing analysis was performed at 6 weeks of life and already showed significant differences in migratory (Agr2,Slit2) and inflammatory (Ccl20, Crispld2) markers between Smad2/3 cKO and control mice. Overall design: Two group comparison: uterine epithelium of control and Smad2/3 cKO mice. We generated a conditional knockout of Smad2/3 in the uterus and demonstrated that Smad2/3 plays a critical role in the endometrium, with disruption resulting in pubertal-onset uterine hyperplasia and ultimately fatal uterine cancer.

在小鼠子宫组织中对Smad2与Smad3进行双条件性敲除（conditional knockout, cKO），可导致子宫内膜失调、不孕以及子宫癌。Smad2/3 cKO小鼠显示出与炎症、细胞周期检验点（cell-cycle checkpoint）、细胞迁移、类固醇生物合成（steroid biosynthesis）以及SMAD1/5调控靶基因相关的基因表达异常。本研究通过RNA测序（RNA-sequencing），鉴定对照组与Smad2/3 cKO小鼠子宫上皮细胞之间的基因表达差异。为控制动情周期（estrous cycle）带来的实验偏差，本研究于交配后0.5天（days post coitum, dpc）收集小鼠子宫上皮组织。本研究对Smad2/3 cKO小鼠与对照组小鼠的全基因组基因表达谱进行了分析。本研究的RNA测序分析于小鼠6周龄时开展，结果已显示Smad2/3 cKO与对照组小鼠在迁移相关标志物（Agr2、Slit2）及炎症相关标志物（Ccl20、Crispld2）上存在显著表达差异。实验整体设计：采用两组对照方案，即对照组与Smad2/3 cKO小鼠的子宫上皮组织。本研究构建了子宫组织特异性Smad2/3条件性敲除小鼠模型，并证实Smad2/3在子宫内膜中发挥关键调控作用，其功能缺失会导致青春期起病的子宫增生症，最终引发致死性子宫癌。