CYP1A1 is overexpressed upon incubation of breast cancer cells with a polyphenolic cocoa extract
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To evaluate the effect of cocoa flavonoids in breast cancer cells at the molecular level, a functional genomic analysis was performed using a polyphenolic cocoa extract (PCE) in MCF-7 and SKBR3 cell lines.
The expression profile of 84 genes included in the Stress & Toxicity PathwayFinder™ PCR Array was analyzed after PCE incubation for 24 h. mRNA and protein levels were analyzed by RT-PCR and western blot, respectively. Gel shift assays were used to evaluate DNA–protein complexes. Protein complexes were identified by co-immunoprecipitation. Cell viability was evaluated by MTT assays.
Upon PCE incubation, 7 genes were overexpressed and 1 underexpressed in MCF-7 cells, whereas 9 genes were overexpressed in SKBR3 cells. Among the differentially expressed genes in both cell lines, cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1) was chosen for further study. CYP1A1 mRNA and protein levels and enzymatic activity increased upon PCE incubation. CYP1A1 transcriptional activation by PCE was mediated through AhR binding to XRE elements within the CYP1A1 promoter in MCF-7 cells. A protein complex including AhR and ERα was detected. The combination of PCE with tamoxifen caused a synergistic cytotoxicity in both cell lines and was due to an increase in apoptosis in MCF-7 cells.
The interaction between ERα and AhR upon incubation with PCE leads to CYP1A1 induction in breast cancer cells. The synergy between PCE and non-cytotoxic tamoxifen concentrations opens the possibility for a combination therapy based on polyphenols from cocoa that increased tamoxifen efficacy.
KeywordsAhR Cocoa CYP1A1 ERα Tamoxifen Breast cancer
Aryl hydrocarbon receptor,
Aryl hydrocarbon receptor nuclear translocator
Bovine serum albumin
Cytochrome P450 family 1 subfamily A polypeptide 1
Electrophoretic mobility shift assay
Polyphenolic cocoa extract
Reverse transcription-polymerase chain reaction
Xenobiotic response element
This work was supported by grants by Nutrexpa SA (CDTI 050618), SAF08-0043 (Ministerio de Educación y Ciencia de España), and ISCIII-RTICc RD06/0020 (Redes Temáticas de Investigación Cooperativa en Salud) RD06/0020/0046. Our research group holds the “quality distinction” from the “Generalitat de Catalunya” SGR2009-00118. CO. was a recipient of a fellowship from the FEC (Federación Española del Café). The authors wish to thank to Dr. Lamuela-Raventós for the analysis of the polyphenol contents in the cocoa samples and Dr. Cascante for her help in the analyses of synergism by statistical methods.
C. Oleaga, M. García, A. Solé, C.J. Ciudad, M. Izquierdo-Pulido, and V. Noé have no conflicts of interest.
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