NUPR1在膀胱癌发展中通过CCR2的转录调控的机械研究
Mechanistic study of NUPR1 in bladder cancer development through transcriptional regulation of CCR2
影响因子:4.00000
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
摘要
核蛋白-1（NUPR1）（也称为P8）是与参与癌症开始和发育各个方面的转录因子相关的基因之一。但是，膀胱癌（BLCA）中NUPR1的分子机制尚不清楚。我们使用基因表达综合（GEO）在线数据库对NUPR1表达与相关基因之间的相关性进行了分析。我们采用了慢病毒介导的小干扰RNA（siRNA）来敲低NUPR1在两个人BLCA细胞系中的表达。进行了体外实验，以验证NUPR1干扰对BLCA的影响以及NUPR1对趋化因子受体-2转录的影响（CCR2）。此外，使用促销数据库预测了CCR2的转录因子。共免疫沉淀（CO-IP）和免疫荧光双染色用于检测NUPR1和CCAAT/增强子结合蛋白γ（CEBPG）之间的结合。进行体内和体外实验，以验证NUPR1通过CEBPG调节CCR2转录。体外实验表明，NUPR1的抑制抑制了BLCA的生长。对GEO数据库的分析表明，NUPR1和CCR2的表达之间存在正相关。荧光素酶实验证实NUPR1会影响CCR2的转录。在线数据表明CEBPG是CCR2的转录因子。 co-IP和免疫荧光双重染色证实了NUPR1和CEBPG之间的结合。荧光素酶测定和染色质免疫沉淀（CHIP）表明CEBPG调节CCR2的转录。此外，在细胞水平上进行的救援实验和动物实验验证了上述机制。 NUPR1在BLCA中促进了促销作用，对NUPR1的干扰可以抑制BLCA的增殖和侵入性。 NUPR1和CCR2的表达之间存在相关性，而NUPR1在细胞核中与CEBPG结合。 NUPR1对CCR2的转录调控可以通过CEBPG参与来实现。
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.