Runt-related transcription factor 1 promotes apoptosis and inhibits neuroblastoma progression in vitro and in vivo
Runt-related transcription factor 1 promotes apoptosis and inhibits neuroblastoma progression in vitro and in vivo
复制标题
Runt 相关转录因子 1 在体外和体内促进细胞凋亡并抑制神经母细胞瘤进展
DOI：
10.1186/s13046-020-01558-2
复制
发表时间：
2020-03-20
影响因子：
11.3
通讯作者：
Li, Shiwang
中科院分区：
文献类型：
--
作者：
Hong, Mei;He, Jing;Li, Shiwang
背景
Runt相关转录因子1（RUNX1）是一种异二聚体转录因子，与许多增强子和启动子的核心元件结合，可以加速多种肿瘤的细胞凋亡。然而，神经母细胞瘤（NB）是一种儿童期高度恶性肿瘤，RUNX1表达的调控机制仍不清楚。在这项研究中，我们旨在评估 RUNX1 在 NB 中的作用，并揭示可能有助于寻找针对 NB 的潜在治疗策略的潜在机制。
方法
使用细胞计数试剂盒-8 (CCK-8) 免疫细胞化学以及涉及软琼脂、细胞侵袭、管形成和整个动物的研究来评估生长、侵袭、转移和血管生成。使用实时定量 PCR 进行 RNA、蛋白质印迹和蛋白质免疫染色分析来测量表达水平。荧光素酶报告基因和染色质免疫沉淀测定表明 RUNX1 直接结合在 BIRC5、CSF2RB 和 NFKBIA 启动子区域内以促进转录。通过测定线粒体膜电位和流式细胞术评估细胞凋亡水平。
结果
相对于低分化（PD）和未分化（UD）组织，RUNX1在神经节神经瘤（GN）和高分化（WD）组织中高表达。此外，RUNX1在体外和体内有效降低细胞活力、侵袭、转移、血管生成并促进细胞凋亡。 RUNX1通过直接结合BIRC5、CSF2RB和NFKBIA启动子来减少BIRC5转录并增加CSF2RB和NFKBIA转录。此外，细胞毒药物，尤其是顺铂，显着增加NB细胞中RUNX1的表达并促进细胞凋亡。
结论
这些数据表明RUNX1是NB进展的独立替代标志物，可用于监测治疗期间的NB预后。
Background
Runt-related transcription factor 1 (RUNX1) is a heterodimeric transcription factor that binds to the core element of many enhancers and promoters and can accelerate apoptosis in various tumors. However, the regulatory mechanisms underlying RUNX1 expression in neuroblastoma (NB), a highly malignant tumor in childhood, remain largely unclear. In this study, we aimed to assess the role of RUNX1 in NB and to reveal the underlying mechanisms that may contribute to finding a potential therapeutics strategy against NB.
Methods
Growth, invasion, metastasis and angiogenesis were assessed using Cell Counting Kit-8 (CCK-8) immunocytochemistry, and studies involving soft agar, cell invasion, tube formation and whole animals. The levels of expression were measured using real-time quantitative PCR for RNA, Western blot and immunostaining analyses for proteins. Luciferase reporter and chromatin immunoprecipitation assays indicated that RUNX1 directly binds within the BIRC5, CSF2RB and NFKBIA promoter regions to facilitate transcription. The level of apoptosis was assessed by determining mitochondrial membrane potential and flow cytometry.
Results
RUNX1 was highly expressed in ganglioneuroma (GN) and well-differentiated (WD) tissues relative to the poorly differentiated (PD) and undifferentiated (UD) ones. Moreover, RUNX1 effectively reduced cell viability, invasion, metastasis, angiogenesis, and promoted apoptosis in vitro and in vivo. RUNX1 reduced BIRC5 transcription and increased CSF2RB and NFKBIA transcription by directly binding BIRC5, CSF2RB and NFKBIA promoters. In addition, cytotoxic drugs, especially cisplatin, significantly increased RUNX1 expression in NB cells and promoted apoptosis.
Conclusions
These data show that RUNX1 is an independent surrogate marker for the progression of NB and it can be used for monitoring NB prognosis during therapy.