ANKHD1 represses p21 (WAF1/CIP1) promoter and promotes multiple myeloma cell growth
ANKHD1 represses p21 (WAF1/CIP1) promoter and promotes multiple myeloma cell growth
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DOI：
10.1016/j.ejca.2014.11.012
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发表时间：
2015-01-01
影响因子：
8.4
通讯作者：
Olalla Saad, Sara T.
中科院分区：
文献类型：
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作者：
Dhyani, Anamika;Machado-Neto, Joao A.;Olalla Saad, Sara T.
ANKHD1（含锚蛋白重复和 KH 结构域的蛋白 1）高表达，在多发性骨髓瘤 (MM) 细胞的增殖和细胞周期进展中发挥重要作用。无论 MM 细胞系的 TP53 突变状态如何，ANKHD1 下调都会调节细胞周期基因表达并上调 p21。本研究旨在探讨ANKHD1在MM体外克隆形成和体内致瘤性中的作用，以及ANKHD1在p21转录调控中的作用。 MM 细胞中 ANKHD1 沉默导致 NO 显着降低。与对照细胞相比，形成的集落数和缓慢迁移（p < 0.05）。此外，在异种移植 MM 小鼠模型中，与对照组相比，注射 ANKHD1 沉默细胞的小鼠肿瘤生长明显受到抑制。与对照组相比，注射沉默细胞的组肿瘤体积（p = 0.006）和重量（p = 0.02）显着减少。免疫共沉淀和染色质免疫沉淀 (ChIP) 测定证实了 p21 和 ANKHD1 之间的相互作用。此外，在荧光素酶检测中，ANKHD1 的过度表达下调了 p21 启动子的活性。荧光素酶活性的降低表明 ANKHD1 在 p21 转录调控中具有直接作用。此外，用 Leptomycin B (LMB) 处理 U266 细胞 24 小时后的共聚焦分析显示，与未处理的细胞相比，ANKHD1 在细胞核内积累，未处理的细胞发现 ANKHD1 主要存在于细胞质中。这表明 ANKHD1 可能在细胞质和细胞核之间穿梭。总之，ANKHD1 通过抑制有效的细胞周期调节因子 p21 来促进 MM 生长。 (C) 2014 Elsevier Ltd. 保留所有权利。
ANKHD1 (Ankyrin repeat and KH domain-containing protein 1) is highly expressed and plays an important role in the proliferation and cell cycle progression of multiple myeloma (MM) cells. ANKHD1 downregulation modulates cell cycle gene expression and upregulates p21 irrespective of the TP53 mutational status of MM cell lines. The present study was aimed to investigate the role of ANKHD1 in MM in vitro clonogenicity and in vivo tumourigenicity, as well as the role of ANKHD1 in p21 transcriptional regulation. ANKHD1 silencing in MM cells resulted in significantly low no. of colonies formed and in slow migration as compared to control cells (p < 0.05). Furthermore, in xenograft MM mice models, tumour growth was visibly suppressed in mice injected with ANKHD1 silenced cells compared to the control group. There was a significant decrease in tumour volume (p = 0.006) as well as in weight (p = 0.02) in the group injected with silenced cells compared to those of the control group. Co-immunoprecipitation and chromatin immunoprecipitation (ChIP) assays confirmed the interaction between p21 and ANKHD1. Moreover, overexpression of ANKHD1 downregulated the activity of a p21 promoter in luciferase assays. Decrease in luciferase activity suggests a direct role of ANKHD1 in p21 transcriptional regulation. In addition confocal analysis after U266 cells were treated with Leptomycin B (LMB) for 24 h showed accumulation of ANKHD1 inside the nucleus as compared to untreated cells where ANKHD1 was found to be predominantly in cytoplasm. This suggests ANKHD1 might be shuttling between cytoplasm and nucleus. In conclusion, ANKHD1 promotes MM growth by repressing p21 a potent cell cycle regulator. (C) 2014 Elsevier Ltd. All rights reserved.