Novel Aryl Hydrocarbon Receptor Agonist Suppresses Migration and Invasion of Breast Cancer Cells

Abstract

BackgroundDespite the remarkable progress to fight against breast cancer, metastasis remains the dominant cause of treatment failure and recurrence. Therefore, control of invasiveness potential of breast cancer cells is crucial. Accumulating evidences suggest Aryl hydrocarbon receptor (Ahr), a helix-loop-helix transcription factor, as a promising target to control migration and invasion in breast cancer cells. Thus, an Ahr-based exploration was performed to identify a new Ahr agonist with inhibitory potentials on cancer cell motility.MethodsFor prediction of potential interactions between Ahr and candidate molecules, bioinformatics analysis was carried out. The interaction of the selected ligand with Ahr and its effects on migration and invasion were examined in vitro using the MDA-MB-231 and T47D cell lines. The silencing RNAs were transfected into cells by electroporation. Expressions of microRNAs (miRNAs) and coding genes were quantified by real-time PCR, and the protein levels were detected by western blot.ResultsThe in silico and in vitro results identified Flavipin as a novel Ahr agonist. It induces formation of Ahr/Ahr nuclear translocator (Arnt) heterodimer to promote the expression of cytochrome P450 family 1 subfamily A member 1 (Cyp1a1). Migration and invasion of MDA-MB-231 and T47D cells were inhibited with Flavipin treatment in an Ahr-dependent fashion. Interestingly, Flavipin suppressed the pro-metastatic factor SRY-related HMG-box4 (Sox4) by inducing miR-212/132 cluster. Moreover, Flavipin inhibited growth and adhesion of both cell lines by suppressing gene expressions of B-cell lymphoma 2 (Bcl2) and integrinα4 (ITGA4).ConclusionTaken together, the results introduce Flavipin as a novel Ahr agonist, and provide first evidences on its inhibitory effects on cancer cell motility, suggesting Flavipin as a candidate to control cell invasiveness in breast cancer patients.

The aryl hydrocarbon receptor (AhR) as a breast cancer drug target

“…The aryl hydrocarbon receptor (AhR) is a member of the basic helix‐loop‐helix/Per‐ARNT‐SIM (bHLH/PAS) transcription factor family, and it has been previously suggested that it is a possible druggable target . It is ligand activated by a variety of synthetic and natural compounds .…”

Characterization of Glyceollins as Novel Aryl Hydrocarbon Receptor Ligands and Their Role in Cell Migration

“…To date, although the role of AhR in cancer remains unclear, and some results are contradictory, many studies have shown the anticancer effects of AhR ligands in breast cancer. Flavipin, an AhR agonist, suppresses the migration and invasion of BC cells [34]. Genistein can confer sensitivity to triple-negative breast cancer for antiestrogen therapy by preventing and reversing AhR-dependent BRCA1 hypermethylation [39].…”

“…Finally, we studied glyceollin effects in cell migration only in the MDA-MB-231 triple-negative breast cancer cells, because cells of this lineage represent the most metastatic type of breast cancer [45]. Moreover, previous studies showed that AhR is an important target of cell migration in triple-negative BC [34,35].…”

“…As several recent studies have indicated that AhR signaling is profoundly involved in cell migration [34,35], we tested the effect of glyceollins on cell migration using an IncuCyte ZOOM ® 96-Well Scratch Wound cell migration assay. We used the triple-negative BC MDA-MB-231 cells, which are known to be highly invasive.…”

Binding of Chlorinated Phenylacrylonitriles to the Aryl Hydrocarbon Receptor: Computational Docking and Molecular Dynamics Simulations

“…The aryl hydrocarbon receptor (AhR) is a member of the basic helix-loop-helix/ Per-ARNT-SIM (bHLH/PAS) transcription factor family [1][2][3][4]. In its inactive state, the AhR resides in the cytosol of the cell as a complex with a number of other proteins.…”