ZNF623 通过招募 CtBP1 来调节 NF-κB 通路促进乳腺癌的进展。

ZNF623 contributes to breast carcinoma progress by recruiting CtBP1 to regulate NF-κB pathway.

机构信息

Immunology Research Center for Oral and Systemic Health, Beijing Friendship Hospital, Capital Medical University, Beijing, 100050, China; Beijing Clinical Research Institute, Beijing, 100050, China.

Medical Research Center, Beijing Institute of Respiratory Medicine and Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China.

出版信息

Biochem Biophys Res Commun. 2024 Oct 8;728:150314. doi: 10.1016/j.bbrc.2024.150314. Epub 2024 Jun 25.

DOI:10.1016/j.bbrc.2024.150314

PMID:38959528

Abstract

BACKGROUND

Breast cancer ranks among the most prevalent tumor types worldwide. Copy number amplification of chromosome 8q24 is frequently detected in breast cancer. ZNF623 is a relatively unexplored gene mapped to 8q24. Here, we explore the expression profile, prognostic significance, and biological action of ZNF623 in breast carcinogenesis.

METHODS

To evaluate the mRNA expression pattern and prognostic relevance of ZNF623 across different cancer types, we conducted bioinformatic analyses. The expression of the gene was suppressed using ZNF623 shRNAs/siRNAs and augmented through transfection with plasmids containing ZNF623 cDNA. Cell viability assay, clonogenic assay, and transwell migration assay were utilized to assess the proliferation, viability, and invasion capacity of breast cancer cell lines. Luciferase reporter assay served as a pivotal tool to ascertain the transcriptional activity of ZNF623. IP-MS and co-IP were employed to validate that ZNF623 interacted with CtBP1. ChIP analysis and ChIP-qPCR were conducted to assess the genes targeted by ZNF623/CtBP1 complex. Flow cytometry was conducted to evaluate the phosphorylation status of p65.

RESULTS

ZNF623 expression was notably elevated in breast cancer (BC). Prognostic analysis indicated higher expression of ZNF623 indicated worse survival. Functional experiments discovered that the upregulation of ZNF623 significantly enhanced both the proliferative and migratory capacities of breast cancer cells. Luciferase reporter assay indicated that ZNF623 was a transcription repressor. Immunoprecipitation coupled mass spectrometry analysis revealed a physical association between ZNF623 and CtBP1 in the interaction group. The conjoint analysis of ChIP-seq and TCGA DEG analysis revealed that the ZNF623/CtBP1 complex repressed a series of genes, such as negative regulation of the NF-kappaB signaling pathway. Flow cytometry analysis discovered that knockdown of ZNF623 decreased the phosphorylation level of p65, indicating that ZNF623 could regulate the activity of the NF-κB pathway.

CONCLUSION

ZNF623 predicts poor prognosis of BC and enhances breast cancer growth and metastasis. By recruiting CtBP1, ZNF623 could suppress NF-κB inhibitors, including COMMD1, NFKBIL1, PYCARD, and BRMS1, expression from the transcription level.

摘要

背景

乳腺癌是全球最常见的肿瘤类型之一。8q24 染色体的拷贝数扩增在乳腺癌中经常被检测到。ZNF623 是一个相对未被探索的基因，位于 8q24。在这里，我们探讨 ZNF623 在乳腺癌发生中的表达谱、预后意义和生物学作用。

方法

为了评估 ZNF623 在不同癌症类型中的 mRNA 表达模式和预后相关性，我们进行了生物信息学分析。使用 ZNF623 shRNAs/siRNAs 抑制基因表达，通过转染含有 ZNF623 cDNA 的质粒增强基因表达。细胞活力测定、集落形成测定和 Transwell 迁移测定用于评估乳腺癌细胞系的增殖、活力和侵袭能力。荧光素酶报告基因测定是确定 ZNF623 转录活性的重要工具。IP-MS 和 co-IP 用于验证 ZNF623 与 CtBP1 相互作用。ChIP 分析和 ChIP-qPCR 用于评估 ZNF623/CtBP1 复合物靶向的基因。流式细胞术用于评估 p65 的磷酸化状态。

结果

ZNF623 在乳腺癌 (BC) 中表达明显上调。预后分析表明，ZNF623 表达水平越高，生存预后越差。功能实验发现，ZNF623 的上调显著增强了乳腺癌细胞的增殖和迁移能力。荧光素酶报告基因测定表明，ZNF623 是一种转录抑制因子。免疫沉淀结合质谱分析显示，在相互作用组中，ZNF623 与 CtBP1 之间存在物理相互作用。ChIP-seq 和 TCGA DEG 分析的联合分析表明，ZNF623/CtBP1 复合物抑制了一系列基因的表达，如 NF-κB 信号通路的负调控。流式细胞术分析发现，ZNF623 的敲低降低了 p65 的磷酸化水平，表明 ZNF623 可以调节 NF-κB 通路的活性。