Chromosomal rearrangements that facilitate tumor formation and progression through activation of oncogenic tyrosine kinases are frequently observed in cancer. The ETV6-NTRK3 (EN) fusion has been implicated in various cancers, including infantile fibrosarcoma, secretory breast carcinoma, and acute myeloblastic leukemia, and has exhibited in vivo and in vitro transforming ability. In the present study, we analyzed transcriptome alterations using DNA microarray and RNA-Seq in EN-transduced NIH3T3 fibroblasts to identify the mechanisms that are involved in EN-mediated tumorigenesis. Through functional profile assessment of EN-regulated transcriptome alterations, we found that upregulated genes by EN were mainly associated with cell motion, membrane invagination, and cell proliferation, while downregulated genes were involved in cell adhesion, which correlated with the transforming potential and increased proliferation in EN-transduced cells. KEGG pathway analysis identified the JAK-STAT signaling pathway with the highest statistical significance. Moreover, Ingenuity Pathway Analysis and gene regulatory network analysis identified the STAT1 transcription factor and its target genes as top EN-regulated molecules. We further demonstrated that EN enhanced STAT1 phosphorylation but attenuated STAT1 acetylation, eventually inhibiting the interaction between the NF-κB p65 subunit and acetylated STAT1. Consequently, nuclear translocation of NF-κB p65 and subsequent NF-κB activity were increased by EN. Notably, inhibition of STAT1 phosphorylation attenuated tumorigenic ability of EN in vitro and in vivo. Taken together, here we report, for the first time, STAT1 as a significant EN-regulated transcription factor and a crucial mediator of EN-induced tumorigenesis.

中文翻译：

---

STAT1作为ETV6-NTRK3诱导的肿瘤发生的关键介质的新型鉴定。

在癌症中经常观察到通过重激活致癌酪氨酸激酶促进染色体形成和进展的染色体重排。ETV6-NTRK3（EN）融合蛋白已涉及多种癌症，包括婴儿纤维肉瘤，分泌性乳腺癌和急性粒细胞性白血病，并已表现出体内和体外转化能力。在本研究中，我们分析了DNA芯片和RNA-Seq在EN转导的NIH3T3成纤维细胞中的转录组改变，以鉴定参与EN介导的肿瘤发生的机制。通过对EN调控的转录组改变的功能概况评估，我们发现EN上调的基因主要与细胞运动，膜内陷和细胞增殖有关，而下调的基因与细胞黏附有关，这与EN转导的细胞中的转化潜力和增殖增加有关。KEGG通路分析确定了具有最高统计学意义的JAK-STAT信号通路。此外，Ingenuity Pathway Analysis和基因调控网络分析将STAT1转录因子及其靶基因确定为EN调控的最高分子。我们进一步证明，EN增强STAT1磷酸化，但减弱STAT1乙酰化，最终抑制NF-κBp65亚基与乙酰化STAT1之间的相互作用。因此，EN增加了NF-κBp65的核易位和随后的NF-κB活性。值得注意的是，STAT1磷酸化的抑制减弱了EN在体外和体内的致瘤能力。总而言之，这是我们第一次报告，