Effect of HDAC3 on promoting the epithelial-mesenchymal transition in triple-negative breast cancer

doi:10.3969/j.issn.1006-7795.2026.04.015

Abstract

Abstract: Objective To investigate the role of histone deacetylase 3 (HDAC3) in the epithelial-mesenchymal transition (EMT) of triple-negative breast cancer (TNBC) cells and elucidate its underlying mechanisms. Methods The expression level of HDAC3 in breast cancer tissues was analyzed based on Gene Expression Omnibus (GEO) database, The Cancer Genome Atlas (TCGA) database and immunohistochemical (IHC) staining. The prognostic potential of HDAC3 expression in TNBC patients was analyzed by using Kaplan-Meier Plotter. Cell proliferation was assessed via growth curve and colony formation assays; Transwell assays were performed to evaluate cell migration and invasion. Western blotting analysis was applied to determine the expression of EMT-related markers. RNA-sequencing (RNA-seq) was performed on MDA-MB-231 cells with HDAC3 knockdown to identify target genes and signaling pathways regulated by HDAC3. Key candidate genes were validated by using reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). Chromatin immunoprecipitation polymerase chain reaction (ChIP-PCR) was used to verify HDAC3 binding to the promoters of target genes. Finally, the expression correlation between HDAC3 and target genes, along with their clinical prognostic significance, were analyzed by using GEO, TCGA databases and Kaplan-Meier Plotter. Results HDAC3 was significantly upregulated in TNBC tissues, and its elevated expression was closely correlated with poor overall survival (OS) and recurrence-free survival (RFS) in patients. Growth curve, colony formation, and Transwell assays demonstrated that HDAC3 overexpression significantly promoted the proliferation, migration, and invasion of TNBC cells. HDAC3 overexpression downregulated the expression of epithelial markers α-catenin and γ-catenin, while upregulating the expression of mesenchymal markers Fibronectin and Vimentin in MDA-MB-231 and SUM 159 cells, whereas HDAC3 knockdown resulted in the opposite effects. RNA-seq analysis identified 693 differentially expressed genes after HDAC3 knockdown, which were enriched in signaling pathways related to malignant tumor progression, such as the PI3K-Akt and NF-κB pathways. RT-qPCR validation confirmed that the expression of genes including acyl-CoA synthetase short-chain family member 1 (ACSS1) and epidermal growth factor (EGF) was downregulated, while that of genes including E-cadherin (CDH1) and superoxide dismutase 2 (SOD2) was upregulated. ChIP-PCR assays confirmed that HDAC3 could specifically bind to the promoter regions of CDH1 and SOD2. Analysis of the GSE31519 dataset showed that HDAC3 expression was significantly negatively correlated with CDH1 and SOD2 expression. Furthermore, Kaplan-Meier survival analysis revealed that low expression of CDH1 and SOD2 was associated with poor prognosis in TNBC patients. Conclusion HDAC3 promotes the proliferation, migration, invasion, and EMT progression of TNBC cells by through transcriptionally repressing the expression of CDH1 and SOD2. These findings suggest that HDAC3 is a potential therapeutic target for TNBC.

Key words: triple-negative breast cancer, histone deacetylase 3, epithelial-mesenchymal transition, cell proliferation, cell migration, cell invasion

CLC Number:

Li Qing#, Wang Siqi#, Ma Tianyu, Ma Xiaoqi, Sun Yuting, Teng Xu, Yu Hefen, Zhang Jingyao△, Huang Wei△. Effect of HDAC3 on promoting the epithelial-mesenchymal transition in triple-negative breast cancer[J]. Journal of Capital Medical University, 2026, 47(4): 738-753.

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