ETV6-NTRK3 fusion oncoprotein transduces NIH 3T3 cells

We report a mouse model that recapitulates expression of the ETV6-NTRK3 (EN) fusion oncoprotein, the product of the t(12;15)(p13;q25) translocation characteristic of human secretory breast carcinoma. Activation of EN expression in mammary tissues by Whey acidic protein (Wap) promoter-driven Cre leads to fully penetrant, multifocal malignant breast cancer with short latency. We provide genetic evidence that committed bipotent or CD61+ luminal alveolar progenitors, are targets of tumorigenesis. Furthermore, EN transforms these otherwise transient progenitors through activation of the AP1 complex. To study the initial effects of ETV6-NTRK3 (EN) mediated transformation, we retrovirally transduced NIH 3T3 cells and generated microarray expression profiling data of EN transduced 3T3 cells as well as control 3T3 cells. Using gene set enrichment analysis (GSEA), we identified a signature involving the AP1 transcriptional complex in EN transduced 3T3 cells. Keywords: genetic modification, cell type comparison We retrovirally transduced NIH 3T3 cells with either EN, or controls (either the empty vector or a kinase-dead version of EN with a mutation at the kinase domain of NTRK3). We then prepared total RNAs from these cells and collected microarray expression profiling data from them using Affymetrix mouse MOE430A chips.

本研究构建了可重现ETV6-NTRK3（EN）融合癌蛋白表达的小鼠模型，该融合癌蛋白是人类分泌性乳腺癌特征性t(12;15)(p13;q25)染色体易位的产物。通过乳清酸性蛋白（Whey Acidic Protein, Wap）启动子驱动的Cre重组酶激活乳腺组织中EN的表达，可诱导完全外显、多灶性的恶性乳腺癌，且潜伏期较短。

本研究提供遗传学证据表明，已定型的双潜能祖细胞或CD61+腔面肺泡祖细胞是肿瘤发生的靶细胞。此外，EN可通过激活AP1复合物，将这些本为一过性的祖细胞转化为肿瘤细胞。

为研究ETV6-NTRK3（EN）介导的转化的初始效应，我们通过逆转录病毒转导NIH 3T3细胞，获取了EN转导组与对照组3T3细胞的微阵列表达谱数据。通过基因集富集分析（Gene Set Enrichment Analysis, GSEA），我们在EN转导的3T3细胞中鉴定出了涉及AP1转录复合物的特征基因集。
关键词：基因修饰，细胞类型比较

我们通过逆转录病毒将EN、对照（空载体或NTRK3激酶结构域突变的激酶失活型EN）转导入NIH 3T3细胞。随后我们从这些细胞中提取总RNA，并使用Affymetrix小鼠MOE430A芯片获取其微阵列表达谱数据。