文章：

ASPP2在有丝分裂期间被CDK1磷酸化，并且是胰腺癌细胞增殖所必需的。

ASPP2 Is Phosphorylated by CDK1 during Mitosis and Required for Pancreatic Cancer Cell Proliferation

原文发布日期：15 November 2023

DOI: 10.3390/cancers15225424

类型: Article

开放获取: 是

英文摘要：

(1) Background: pancreatic cancer is highly lethal. The role of apoptosis-stimulating protein of p53-2 (ASPP2) in this lethal disease remains unclear. This protein belongs to the ASPP family of p53 interacting proteins. Previous studies in this lab used phosphate-binding tag (Phos-tag) sodium dodecyl sulfate (SDS) polyacrylamide gels and identified a motility upshift of the ASPP family of proteins during mitosis. (2) Purpose: this study expands on previous findings to identify the detailed phosphorylation regulation of ASPP2 during mitosis, as well as the function of ASPP2 in pancreatic cancer. (3) Methods: the Phos-tag technique was used to investigate the phosphorylation mechanism of ASPP2 during mitosis. Phospho-specific antibodies were generated to validate the phosphorylation of ASPP2, and ASPP2-inducible expression cell lines were established to determine the role of ASPP2 in pancreatic cancer. RNA sequencing (RNA-Seq) was used to uncover the downstream targets of ASPP2. (4) Results: results demonstrate that ASPP2 is phosphorylated during mitosis by cyclin-dependent kinase 1 (CDK1) at sites S562 and S704. In vitro and in vivo results show that ASPP2 is required for pancreatic cancer growth. Furthermore, the expressions of yes-associated protein (YAP)-related genes are found to be dramatically altered by ASPP2 depletion. Together, these findings reveal the phosphorylation mechanism of ASPP2 during mitosis. Collectively, results strongly indicate that ASPP2 is a potential target for abating tumor cell growth in pancreatic cancer.

摘要翻译： 

（1）背景：胰腺癌具有极高的致死率。p53凋亡刺激蛋白2（ASPP2）在这一致命疾病中的作用尚不明确。该蛋白属于p53相互作用蛋白ASPP家族。本实验室前期研究采用磷酸结合标签（Phos-tag）十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶电泳技术，发现ASPP家族蛋白在有丝分裂期呈现迁移率上移现象。（2）目的：本研究在前期发现基础上，旨在阐明ASPP2在有丝分裂期的具体磷酸化调控机制及其在胰腺癌中的功能。（3）方法：采用Phos-tag技术探究ASPP2在有丝分裂期的磷酸化机制；制备磷酸化特异性抗体验证ASPP2磷酸化状态；建立ASPP2诱导表达细胞系以确定其在胰腺癌中的作用；通过RNA测序技术揭示ASPP2的下游靶标。（4）结果：研究证实ASPP2在有丝分裂期被细胞周期蛋白依赖性激酶1（CDK1）在S562和S704位点磷酸化。体外及体内实验表明ASPP2为胰腺癌细胞生长所必需。此外，研究发现ASPP2缺失会显著改变Yes相关蛋白（YAP）相关基因的表达。这些发现共同揭示了ASPP2在有丝分裂期的磷酸化机制，充分表明ASPP2是抑制胰腺癌细胞生长的潜在治疗靶点。

原文链接：

ASPP2 Is Phosphorylated by CDK1 during Mitosis and Required for Pancreatic Cancer Cell Proliferation