Lysine-specific demethylase 1 (LSD1), a histone lysine demethylase with the main specificity for H3K4me2, has been shown to be overexpressed in rhabdomyosarcoma (RMS) tumor samples. However, its role in RMS biology is not yet well understood. Here, we identified a new role of LSD1 in regulating adhesion of RMS cells. Genetic knockdown of LSD1 profoundly suppressed clonogenic growth in a panel of RMS cell lines, whereas LSD1 proved to be largely dispensable for regulating cell death and short-term survival. Combined RNA and ChIP-sequencing performed to analyze RNA expression and histone methylation at promoter regions revealed a gene set enrichment for adhesion-associated terms upon LSD1 knockdown. Consistently, LSD1 knockdown significantly reduced adhesion to untreated surfaces. Importantly, precoating of the plates with the adhesives collagen I or fibronectin rescued this reduced adhesion of LSD1 knockdown cells back to levels of control cells. Using KEGG pathway analysis, we identified 17 differentially expressed genes (DEGs) in LSD1 knockdown cells related to adhesion processes, which were validated by qRT-PCR. Combining RNA and ChIP-sequencing results revealed that, within this set of genes, SPP1, C3AR1, ITGA10 and SERPINE1 also exhibited increased H3K4me2 levels at their promoter regions in LSD1 knockdown compared to control cells. Indeed, LSD1 ChIP experiments confirmed enrichment of LSD1 at their promoter regions, suggesting a direct transcriptional regulation by LSD1. By identifying a new role of LSD1 in the modulation of cell adhesion and clonogenic growth of RMS cells, these findings highlight the importance of LSD1 in RMS.

中文翻译：

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下一代测序揭示了赖氨酸特异性脱甲基酶1在横纹肌肉瘤细胞粘附中的新作用。

赖氨酸特异性脱甲基酶1（LSD1），一种对H3K4me2具有主要特异性的组蛋白赖氨酸脱甲基酶，已被证明在横纹肌肉瘤（RMS）肿瘤样品中过表达。但是，其在RMS生物学中的作用尚未得到很好的理解。在这里，我们确定了LSD1在调节RMS细胞粘附中的新作用。LSD1的基因敲低极大地抑制了一组RMS细胞系中克隆形成的生长，而LSD1被证明在调节细胞死亡和短期存活方面是主要可有可无的。结合RNA和ChIP测序分析启动子区域的RNA表达和组蛋白甲基化，揭示了LSD1敲除后与粘附相关的术语的基因集富集。始终如一地，LSD1组合式大大降低了对未处理表面的附着力。重要的，用胶粘剂I或纤连蛋白对板进行预涂可以挽救LSD1敲低细胞的这种减少的粘附力，使其回到对照细胞的水平。使用KEGG通路分析，我们在与粘附过程相关的LSD1敲低细胞中鉴定了17个差异表达基因（DEG），这些基因已通过qRT-PCR验证。结合RNA和ChIP测序的结果表明，在这组基因中，与对照细胞相比，SSD1，C3AR1，ITGA10和SERPINE1在LSD1敲低的启动子区域也表现出增加的H3K4me2水平。确实，LSD1 ChIP实验证实了LSD1在其启动子区域的富集，表明LSD1可以直接进行转录调控。通过发现LSD1在调节RMS细胞的细胞粘附和克隆生长中的新作用，这些发现凸显了LSD1在RMS中的重要性。