Objective:
Triple-negative breast cancer (TNBC) is a subtype of breast cancer that has a worse prognosis than the other subtypes of breast cancer because of its high recurrence and metastasis rates. The objective of this study is to identify the regulatory factors that are associated with the disease-free survival (DFS) of TNBC and potential biomarkers for TNBC treatment.
Material and Methods:
We obtained the GSE97342 dataset from the Gene Expression Omnibus website and conducted weighted gene co-expression network analysis (WGCNA) to identify modules associated with the DFS of TNBC. Subsequently, biological functions of the modules were elucidated through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. Cross-checking with the Human Transcription Factor Database facilitated the selection of hub transcription factors through univariate Cox regression analysis of overlapping transcription factors. Utilizing bioinformatics analysis, we assessed the prognostic significance of these hub transcription factors, investigated their target genes, and explored their associations with tumor immune cells in TNBC. Finally, the expression levels of the hub transcription factors were validated by immunohistochemical staining, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and Western blotting.
Results:
Through WGCNA analysis, we identified three modules correlated with DFS in TNBC. GO and KEGG analyses elucidated the biological functions of genes within these modules. Survival analysis pinpointed three hub transcription factors: Forkhead box D1 (FOXD1), aryl hydrocarbon receptor nuclear translocator 2 (ARNT2), and zinc finger protein 132 (ZNF132). The expression level of FOXD1 was negatively associated with the prognoses of patients with TNBC, whereas the other two genes were positively associated with the prognoses of patients with TNBC. Immunohistochemical staining, qRT-PCR, and Western blotting validated the expression levels of the hub transcription factors.
Conclusion:
We discovered three hub transcription factors (FOXD1, ARNT2, and ZNF132) that were correlated with the DFS of TNBC. These correlations suggested their potential as prognostic predictors for patients with TNBC.
