Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells.
Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells.
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RPS27a 表达缺失通过 RPL11-MDM2-p53 通路调节肺腺癌细胞的细胞周期、凋亡和增殖
DOI：
10.1186/s13046-021-02230-z
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发表时间：
2022-01-24
期刊：
影响因子：
--
通讯作者：
An X
中科院分区：
文献类型：
--
作者：
Li H;Zhang H;Huang G;Bing Z;Xu D;Liu J;Luo H;An X
某些核糖体蛋白的消耗诱导p53活化，其主要由核糖体蛋白L5（RPL 5）和/或核糖体蛋白L11（RPL 11）介导。因此，RPL 5和RPL 11可能将RP和p53激活联系起来。因此，本研究旨在探讨RP是否与RPL 11相互作用并调节肺腺癌（LUAD）细胞中p53的活化。通过免疫沉淀法将A549细胞中的内源性RPL 11结合蛋白拉下，并通过蛋白质组学方法鉴定。对接分析和GST融合蛋白分析用于分析核糖体蛋白S27 a（RPS 27 a）和RPL 11的相互作用。采用免疫共沉淀和体外泛素化实验检测RPS 27 a基因敲减对RPS 27 a与RPL 11相互作用以及对p53积累的影响。在RPS 27 a敲低的存在下进行细胞周期、细胞凋亡、细胞侵袭和迁移、细胞活力和集落形成测定。根据TCGA数据集分析LUAD中RPS 27 a mRNA的表达，并通过免疫组化检测LUAD样本中RPS 27 a的表达。最后，通过免疫组织化学在RPS 27 a敲低的A549细胞异种移植物中分析RPS 27 a和p53表达。RPS 27 a是一种新的RPL 11结合蛋白。GST pull-down分析显示RPS 27 a直接结合RPL 11。RPS 27 a的敲低减弱了RPS 27 a与RPL 11之间的相互作用，但增强了RPL 11与小鼠双微体2（MDM 2）的结合，从而抑制了MDM 2对p53的泛素化和降解。RPS 27 a的敲除以RPL 11依赖的方式稳定p53，并以p53依赖的方式在A549细胞中诱导细胞活力抑制、细胞周期停滞和凋亡。RPS 27 a在LUAD中表达上调，且与LUAD的进展和预后相关。在LUAD临床标本中，RPS 27 a的过表达与p53、MDM 2和RPL 11的上调相关。RPS 27 a的敲低增加了p53活化，因此抑制了裸鼠中A549细胞异种移植物的形成。RPS 27 a与RPL 11相互作用，RPS 27 a的敲低可增强RPL 11与MDM 2的结合，从而抑制MDM 2介导的p53泛素化和降解;此外，RPS 27 a在LUAD的进展和预后中起重要作用，可能成为LUAD患者的治疗靶点。在线版本包含补充材料，可通过10.1186/s13046-021-02230-z获得。
Depletion of certain ribosomal proteins induces p53 activation, which is mediated mainly by ribosomal protein L5 (RPL5) and/or ribosomal protein L11 (RPL11). Therefore, RPL5 and RPL11 may link RPs and p53 activation. Thus, this study aimed to explore whether RPs interact with RPL11 and regulate p53 activation in lung adenocarcinoma (LUAD) cells. The endogenous RPL11-binding proteins in A549 cells were pulled down through immunoprecipitation and identified with a proteomics approach. Docking analysis and GST-fusion protein assays were used to analyze the interaction of ribosomal protein S27a (RPS27a) and RPL11. Co-immunoprecipitation and in vitro ubiquitination assays were used to detect the effects of knockdown of RPS27a on the interaction between RPS27a and RPL11, and on p53 accumulation. Cell cycle, apoptosis, cell invasion and migration, cell viability and colony-formation assays were performed in the presence of knockdown of RPS27a. The RPS27a mRNA expression in LUAD was analyzed on the basis of the TCGA dataset, and RPS27a expression was detected through immunohistochemistry in LUAD samples. Finally, RPS27a and p53 expression was analyzed through immunohistochemistry in A549 cell xenografts with knockdown of RPS27a. RPS27a was identified as a novel RPL11 binding protein. GST pull-down assays revealed that RPS27a directly bound RPL11. Knockdown of RPS27a weakened the interaction between RPS27a and RPL11, but enhanced the binding of RPL11 and murine double minute 2 (MDM2), thereby inhibiting the ubiquitination and degradation of p53 by MDM2. Knockdown of RPS27a stabilized p53 in an RPL11-dependent manner and induced cell viability inhibition, cell cycle arrest and apoptosis in a p53-dependent manner in A549 cells. The expression of RPS27a was upregulated in LUAD and correlated with LUAD progression and poorer prognosis. Overexpression of RPS27a correlated with upregulation of p53, MDM2 and RPL11 in LUAD clinical specimens. Knockdown of RPS27a increased p53 activation, thus, suppressing the formation of A549 cell xenografts in nude mice. RPS27a interacts with RPL11, and RPS27a knockdown enhanced the binding of RPL11 and MDM2, thereby inhibiting MDM2-mediated p53 ubiquitination and degradation; in addition, RPS27a as important roles in LUAD progression and prognosis, and may be a therapeutic target for patients with LUAD. The online version contains supplementary material available at 10.1186/s13046-021-02230-z.
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DOI：
10.4161/nucl.32235
发表时间：
2014-09
期刊：
Nucleus (Austin, Tex.)
影响因子：
--
作者：
James A;Wang Y;Raje H;Rosby R;DiMario P
通讯作者：
DiMario P
DOI：
10.1186/s13046-018-0956-3
发表时间：
2018-11-26
期刊：
Journal of experimental & clinical cancer research : CR
影响因子：
--
作者：
Zhou S;Lu J;Li Y;Chen C;Cai Y;Tan G;Peng Z;Zhang Z;Dong Z;Kang T;Tang F
通讯作者：
Tang F
DOI：
10.1016/j.bbrc.2017.07.105
发表时间：
2017-09-16
影响因子：
3.1
作者：
Hong, Seung-Woo;Kim, Seung-Mi;Hur, Dae Young
通讯作者：
Hur, Dae Young
影响因子：
5.4
作者：
Deisenroth C;Franklin DA;Zhang Y
通讯作者：
Zhang Y
核糖体蛋白 S26 调节 p53 活性以应对 DNA 损伤
影响因子：
8
作者：
Cui, D.;Li, L.;Tang, J.
通讯作者：
Tang, J.