Nfatc1 enhances stem cell contribution to squamous skin tumors. Mus musculus

We find that epithelial-specific deletion of Nfatc1 decreases skin tumor susceptibility in mice subjected to DMBA/TPA carcinogenesis. To probe the molecular mechanisms by which Nfatc1 may regulate tumorigenesis, we identify gene expression changes associated with constitutive NFATc1 activation in primary mouse keratinocytes. Constitutively active NFATc1 activity in primary mouse keratinocytes to investigate how Nfatc1 regulates tumor initiation in the skin. Overall design: Primary mouse keratinocytes were infected with retroviruses encoding a recombinant human NFATc1 containing mutations in the autoinhibitory domain that render it constitutively active (CAC1). Keratinocytes infected with retroviruses generated from an empty MSCV reotroviral vector were used as a control. Three biological replicates were performed.

我们发现，在经DMBA/TPA致癌方案处理的小鼠中，上皮特异性敲除核因子活化T细胞c1（Nfatc1）可降低皮肤肿瘤易感性。为探究Nfatc1调控肿瘤发生的分子机制，我们在原代小鼠角质形成细胞中鉴定出与组成型Nfatc1活化相关的基因表达变化。本研究通过将组成型活化的Nfatc1导入原代小鼠角质形成细胞，以解析Nfatc1如何调控皮肤肿瘤起始过程。整体实验设计：将编码重组人Nfatc1的逆转录病毒感染原代小鼠角质形成细胞，该重组蛋白的自抑制结构域携带突变，可使其呈现组成型活化状态（CAC1）；以空MSCV逆转录病毒载体包装的逆转录病毒感染的角质形成细胞作为对照。本实验共设置三次生物学重复。