Characterization and Cytotoxicity of West Java Coffee Extracts, Chlorogenic Acid, Caffeine on MDA-MB-231, MCF-7 Cell Lines, and Their Impact on MYC and AXIN2 Wnt Pathway Gene Expression
Abstract
Background: Coffee bean extracts have been widely investigated for their potential anticancer properties, particularly due to bioactive compounds such as chlorogenic acid and caffeine. Breast cancer development involves complex genetic and signaling alterations, including dysregulation of the Wnt/β-catenin pathway.
Purpose: To determine the phytochemical composition and quantify caffeine and chlorogenic acid in West Java coffee bean extracts. The study assessed their effects on the viability of MDA-MB-231 and MCF-7 breast cancer cells, and examined impacts on the Wnt pathway, focusing on MYC and AXIN2 expression.
Methods: Phytochemical screening and HPLC analysis are used to measure chlorogenic acid and caffeine in green and roasted coffee extracts. Cytotoxic activity and IC5 0 values in MDA-MB-231 and MCF-7 breast cancer cells were assessed with the WST-8 assay. Gene expression of MYC and AXIN2 was analyzed by q-RT-PCR to assess modulation of the Wnt pathway. Cytotoxic selectivity was tested in HEK293 cells. Flow cytometry analyzes apoptosis and the cell cycle. Molecular mechanisms were explored using docking, 200-ns molecular dynamics simulations, and MM-PBSA binding free energy calculations targeting Tankyrase-1 and Estrogen Receptor Alpha.
Results: Roasting and extraction methods affected phytochemicals, with higher temperatures reducing chlorogenic acid levels. Coffee extracts showed cytotoxicity against MDA-MB-231 (IC5 0 258.4-637 µg/mL) and MCF-7 (IC5 0 190.7-686…
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