State Key Laboratory of Medical Genomics, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
Medical Institute of Health Sciences, Chinese Academy of Sciences, Shanghai, 200025, China.
Front Med. 2017 Mar;11(1):87-96. doi: 10.1007/s11684-016-0469-4. Epub 2017 Mar 2.
S100A9, a calcium-binding protein, participates in the inflammatory process and development of various tumors, thus attracting much attention in the field of cancer biology. This study aimed to investigate the regulatory mechanism of S100A9 and its function involvement in APL. We used real-time quantitative PCR to determine whether PML/RARα affects the expression of S100A9 in NB4 and PR9 cells upon ATRA treatment. ChIP-based PCR and dual-luciferase reporter assay system were used to detect how PML/RARα and PU.1 regulate S100A9 promoter activity. CCK-8 assay and flow cytometry were employed to observe the viability and apoptosis of NB4 cells when S100A9 was overexpressed. Results showed that S100A9 was an ATRA-responsive gene, and PML/RARα was necessary for the ATRA-induced expression of S100A9 in APL cells. In addition, PU.1 could bind to the promoter of S100A9, especially when treated with ATRA in NB4 cells, and promote its activity. More importantly, overexpression of S100A9 induced the apoptosis of NB4 cells and inhibited cell growth. Collectively, our data indicated that PML/RARα and PU.1 were necessary for the ATRA-induced expression of S100A9 in APL cells. Furthermore, S100A9 promoted apoptosis in APL cells and affected cell growth.
S100A9 是一种钙结合蛋白，参与多种肿瘤的炎症过程和发展，因此在癌症生物学领域引起了广泛关注。本研究旨在探讨 S100A9 的调控机制及其在 APL 中的功能作用。我们使用实时定量 PCR 来确定 PML/RARα 是否会影响 ATRA 处理后 NB4 和 PR9 细胞中 S100A9 的表达。使用 ChIP 基于 PCR 和双荧光素酶报告基因检测系统来检测 PML/RARα 和 PU.1 如何调节 S100A9 启动子活性。使用 CCK-8 检测试剂盒和流式细胞术来观察 NB4 细胞中 S100A9 过表达时的细胞活力和凋亡情况。结果表明，S100A9 是 ATRA 反应性基因，PML/RARα 是 APL 细胞中 ATRA 诱导 S100A9 表达所必需的。此外，PU.1 可以结合 S100A9 启动子，尤其是在 NB4 细胞中用 ATRA 处理时，可促进其活性。更重要的是，S100A9 的过表达诱导 NB4 细胞凋亡并抑制细胞生长。综上所述，我们的数据表明 PML/RARα 和 PU.1 是 APL 细胞中 ATRA 诱导 S100A9 表达所必需的。此外，S100A9 促进 APL 细胞凋亡并影响细胞生长。
