Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Università Federico II, 80131 Napoli, Italy.
Dipartimento di Medicina Sperimentale e Clinica, Università degli Studi di Firenze, viale Morgagni 50, 50134 Firenze, Italy.
Nucleic Acids Res. 2020 Sep 18;48(16):8943-8958. doi: 10.1093/nar/gkaa599.
The epithelial-to-mesenchymal transition (EMT) is a complex transcriptional program induced by transforming growth factor β1 (TGF-β1). Histone lysine-specific demethylase 1 (LSD1) has been recognized as a key mediator of EMT in cancer cells, but the precise mechanism that underlies the activation and repression of EMT genes still remains elusive. Here, we characterized the early events induced by TGF-β1 during EMT initiation and establishment. TGF-β1 triggered, 30-90 min post-treatment, a nuclear oxidative wave throughout the genome, documented by confocal microscopy and mass spectrometry, mediated by LSD1. LSD1 was recruited with phosphorylated SMAD2/3 to the promoters of prototypic genes activated and repressed by TGF-β1. After 90 min, phospho-SMAD2/3 downregulation reduced the complex and LSD1 was then recruited with the newly synthesized SNAI1 and repressors, NCoR1 and HDAC3, to the promoters of TGF-β1-repressed genes such as the Wnt soluble inhibitor factor 1 gene (WIF1), a change that induced a late oxidative burst. However, TGF-β1 early (90 min) repression of transcription also required synchronous signaling by reactive oxygen species and the stress-activated kinase c-Jun N-terminal kinase. These data elucidate the early events elicited by TGF-β1 and the priming role of DNA oxidation that marks TGF-β1-induced and -repressed genes involved in the EMT.
上皮间质转化（EMT）是转化生长因子β1（TGF-β1）诱导的复杂转录程序。组蛋白赖氨酸特异性去甲基化酶 1（LSD1）已被认为是癌细胞 EMT 的关键介质，但 EMT 基因激活和抑制的确切机制仍不清楚。在这里，我们描述了 TGF-β1 在 EMT 起始和建立过程中诱导的早期事件。TGF-β1 在处理后 30-90 分钟内触发了整个基因组的核氧化波，这通过共聚焦显微镜和质谱法得到了证实，由 LSD1 介导。LSD1 与磷酸化 SMAD2/3 一起被募集到 TGF-β1 激活和抑制的原型基因的启动子上。90 分钟后，磷酸化 SMAD2/3 的下调减少了复合物，然后 LSD1 与新合成的 SNAI1 和抑制剂 NCoR1 和 HDAC3 一起被募集到 TGF-β1 抑制基因的启动子上，如 Wnt 可溶性抑制剂因子 1 基因（WIF1），这种变化诱导了晚期氧化爆发。然而，TGF-β1 早期（90 分钟）对转录的抑制也需要活性氧和应激激活激酶 c-Jun N 端激酶的同步信号转导。这些数据阐明了 TGF-β1 引发的早期事件以及 DNA 氧化的启动作用，标志着 EMT 中涉及的 TGF-β1 诱导和抑制的基因。
