NUPR1在膀胱癌发生中的机制研究：转录调控CCR2

Mechanistic study of NUPR1 in bladder cancer development through transcriptional regulation of CCR2

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JOURNAL OF CELLULAR PHYSIOLOGY

影响因子:4

分区:生物学3区 / 细胞生物学3区生理学3区

发表日期:2024 Nov

作者: Zebin Shi, Yuanyuan Mi, Li Zhang, Wenxu Zhang, Wei Zhang, Xiaokai Shi, Shenglin Gao, Li Zuo, Lifeng Zhang

DOI: 10.1002/jcp.31412

摘要

核蛋白-1（NUPR1）（也称为p8）是与转录因子相关的基因之一，参与癌症的多方面发生和发展。然而，NUPR1在膀胱癌（BLCA）中的分子机制尚不清楚。我们利用Gene Expression Omnibus（GEO）数据库分析了NUPR1表达与相关基因的相关性。采用慢病毒介导的小干扰RNA（siRNA）敲低两个人类膀胱癌细胞系中的NUPR1表达。在体外实验中验证了NUPR1干扰对膀胱癌的影响及NUPR1对趋化因子受体2（CCR2）转录的影响。此外，利用PROMO数据库预测CCR2的转录因子。采用免疫共沉淀（Co-IP）和免疫荧光双染色检测NUPR1与CCAAT/增强子结合蛋白γ（CEBPG）之间的结合。在体内和体外实验中验证NUPR1通过CEBPG调控CCR2转录的机制。体外实验显示，抑制NUPR1可以抑制膀胱癌的生长。GEO数据库分析表明NUPR1与CCR2的表达呈正相关。荧光素酶实验证实NUPR1影响CCR2的转录。在线数据库显示，CEBPG是CCR2的转录因子。Co-IP和免疫荧光双染色确认NUPR1与CEBPG的结合。荧光素酶检测和染色质免疫沉淀（ChIP）表明CEBPG调控CCR2的转录。此外，细胞水平的拯救实验和动物实验验证了上述机制。NUPR1在膀胱癌中具有促进作用，干扰NUPR1可以抑制膀胱癌的增殖和侵袭能力。NUPR1与CCR2的表达相关，并在细胞核中与CEBPG结合，NUPR1通过CEBPG参与调控CCR2的转录。

Abstract

Nuclear protein-1 (NUPR1) (also known as p8) is one of the genes associated with transcription factors that participate in various aspects of cancer initiation and development. However, the molecular mechanisms of NUPR1 in bladder cancer (BLCA) remain unclear. We conducted an analysis of the correlation between NUPR1 expression and related genes using the Gene Expression Omnibus (GEO) online database. We employed lentivirus-mediated small interfering RNA (siRNA) to knockdown the expression of NUPR1 in two human BLCA cell lines. In vitro experiments were conducted to validate the impact of NUPR1 interference on BLCA and the influence of NUPR1 on the transcription of chemokine receptor-2 (CCR2). Furthermore, transcription factors for CCR2 were predicted using the PROMO database. Co-immunoprecipitation (Co-IP) and immunofluorescence double staining were used to detect the binding between NUPR1 and CCAAT/enhancer binding protein γ (CEBPG). In vivo and in vitro experiments were conducted to validate that NUPR1 regulates CCR2 transcription through CEBPG. In vitro experiments indicate that the suppression of NUPR1 inhibited BLCA growth. Analysis of the GEO database revealed a positive correlation between the expression of NUPR1 and CCR2. Luciferase experiments confirmed that NUPR1 influences the transcription of CCR2. Online data indicates that CEBPG is a transcription factor for CCR2. Co-IP and immunofluorescence double staining confirmed binding between NUPR1 and CEBPG. Luciferase assays and chromatin immunoprecipitation (ChIP) demonstrate that CEBPG regulates the transcription of CCR2. Additionally, rescue experiments at the cellular level and animal experiments validated the aforementioned mechanism. NUPR1 promotes a promotional role in BLCA, and interference with NUPR1 can inhibit the proliferation and invasive abilities of BLCA. There was a correlation between the expressions of NUPR1 and CCR2, and NUPR1 binds with CEBPG in the cell nucleus. Transcriptional regulation of CCR2 by NUPR1 may be achieved through the involvement of CEBPG.