Induction of JUN and FOS Expression by Gallic Acid Derivatives in MCF-7 Breast Cancer Cells: An in Silico and RT-PCR Study
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Abstract
Breast cancer remains the leading malignancy worldwide and continues to cause high morbidity and mortality. Although chemotherapy, hormonal therapy, immunotherapy, and targeted treatments are available, these therapeutic options frequently induce adverse effects and contribute to resistance, limiting long-term outcomes. Gallic acid and its derivatives have been widely reported to exert cytotoxic and pro-apoptotic effects on various cancer cell lines, including MCF7 breast cancer cells. However, the underlying molecular mechanisms, particularly those involving apoptosis-related gene modulation, have not been fully elucidated. This study investigated the expression changes of key apoptosis-related genes in MCF7 cells following treatment with two gallic acid derivatives, N-octyl gallamide and N-tert-butyl gallamide, using integrated in silico and in vitro approaches. Differentially expressed genes (DEGs) were identified from the GSE158788 dataset using GEO2R. Protein–protein interaction networks were constructed using Cytoscape, and KEGG pathway enrichment was performed to determine relevant signaling pathways. JUN and FOS, identified as hub genes, were validated using quantitative RT-PCR in MCF7 cells treated with IC₅₀ and 2×IC₅₀ concentrations of the test compounds. DEG and PPI analyses identified JUN and FOS as key hub genes associated with gallic acid–mediated apoptosis. RT-PCR results demonstrated that N-octyl gallamide significantly upregulated JUN (ΔΔCTs 1.25 ± 0.251; p < 0.05) and FOS (1.82 ± 0.691; p < 0.05), exceeding the effects of tamoxifen. In contrast, N-tert-butyl gallamide did not significantly alter JUN or FOS expression (ΔΔCTs 0.76 ± 0.053; p > 0.05). N-octyl gallamide exhibits strong potential as an anticancer agent through JUN and FOS activation in MCF7 cells. These findings highlight its promise for further development as an alternative or adjuvant breast cancer therapy.
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