Department of Gastrointestinal Surgery, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, P.R. China.
Department of Microbiology and Immunology, Institute of Molecular Virology and Immunology, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang Province, P.R. China.
Cell Death Dis. 2020 Jul 8;11(7):514. doi: 10.1038/s41419-020-2718-3.
Death domain-associated protein (DAXX) is a complex biological multifunctional protein and is involved in the tumorigenesis and progression of multiple cancers. The accumulation of DAXX in the nucleus is a common phenomenon in tumor cells. However, altering the subcellular localizations of DAXX results in different biological functions, and we also found that its nuclear/cytoplasmic ratio (NCR) was associated with poor prognosis in gastric cancer (GC). In this study, we investigated the effect of cytoplasmic and nuclear DAXX (cDAXX and nDAXX) in GC and the underlying mechanisms. Immunohistochemical detection performed in 323 GC tissues reveled that cDAXX was associated with a better survival, while high nDAXX expression suggested a poorer prognosis outcome. Upregulation of DAXX in the cytoplasm inhibited cell proliferation and promoted apoptosis, whereas downregulation of DAXX in the nucleus displayed opposite effects. Moreover, Transwell assays revealed that DAXX enhanced GC cell migration and invasion. Analysis from the Gene Expression Profile Interactive Analysis (GEPIA) database showed that the expression of DAXX was significantly associated with SUMO-2/3 in GC tissues. Co-immunoprecipitation combined with immunofluorescence analysis indicated that DAXX interacted directly with SUMO-2/3. Subsequently, down-regulating the expression of SUMO-2/3 resulted in altered subcellular localization of DAXX. Bioinformatics analysis showed that RanBP2 may act as SUMO E3 ligase to promote nuclear-plasma transport via combining with RanGAP1. Taken together, our results indicated that DAXX plays opposing roles in GC and suggest a new model whereby cDAXX, nDAXX, and SUMO-2/3 form a molecular network that regulates the subcellular localization of DAXX and thereby modulates its opposing biological effects. Thus, our findings provide a foundation for future studies of DAXX as a novel therapeutic target for patients with GC.
死亡结构域相关蛋白(DAXX)是一种复杂的生物多功能蛋白,参与多种癌症的发生和发展。DAXX 在细胞核中的积累是肿瘤细胞的一个常见现象。然而,改变 DAXX 的亚细胞定位会导致不同的生物学功能,我们还发现其核/细胞质比(NCR)与胃癌(GC)的预后不良有关。在这项研究中,我们研究了 GC 中细胞质和核内 DAXX(cDAXX 和 nDAXX)的作用及其潜在机制。在 323 份 GC 组织中进行的免疫组织化学检测表明,cDAXX 与更好的生存相关,而高 nDAXX 表达提示预后不良。细胞质中 DAXX 的上调抑制细胞增殖并促进细胞凋亡,而核内 DAXX 的下调则显示出相反的效果。此外,Transwell 测定表明 DAXX 增强了 GC 细胞的迁移和侵袭。从基因表达谱交互分析(GEPIA)数据库的分析表明,DAXX 在 GC 组织中的表达与 SUMO-2/3 显著相关。免疫共沉淀结合免疫荧光分析表明,DAXX 与 SUMO-2/3 直接相互作用。随后,下调 SUMO-2/3 的表达导致 DAXX 的亚细胞定位发生改变。生物信息学分析表明,RanBP2 可能作为 SUMO E3 连接酶通过与 RanGAP1 结合来促进核浆转运。总之,我们的研究结果表明,DAXX 在 GC 中发挥着相反的作用,并提出了一个新的模型,即 cDAXX、nDAXX 和 SUMO-2/3 形成一个分子网络,调节 DAXX 的亚细胞定位,从而调节其相反的生物学效应。因此,我们的研究结果为 DAXX 作为 GC 患者的新型治疗靶点的未来研究提供了基础。