3-8-dihydroxy-6h-dibenzo(b-d)pyran-6-one and Breast-Neoplasms

While preclinical studies suggest the breast cancer (BC) chemopreventive effects of dietary polyphenols, the human evidence is still very weak. The huge existing in vitro-in vivo gap is mainly due to the plethora of potential effects reported by in vitro studies that usually assay polyphenols as occurring in the food (beverages, extracts, foods) and/or derived aglycone metabolites with doubtful physiological relevance. Since phase-II metabolites can reach systemic tissues such as malignant breast tumors, we aimed here to compare for the first time the antiproliferative and estrogenic/antiestrogenic effects of dietary polyphenols and microbiota-derived metabolites (i.e., resveratrol, dihydroresveratrol, urolithins (A, B, and Isourolithin A), and the flavanone hesperetin), with those effects exerted by their physiologically relevant glucuronides and sulfates on human BC cell lines (MDA-MB-231 and MCF-7). Results showed that aglycones exerted dose-dependent antiproliferative and estrogenic/antiestrogenic activities, but both their glucuronide and sulfate conjugates lacked these activities. In addition, aglycones underwent phase-II metabolism in BC cells, mainly via sulfation, which determined the cell-dependent differences in the effects observed. Therefore, phase-II metabolism limits the antiproliferative, estrogenic, and antiestrogenic activities of dietary polyphenols on BC cells. Likewise, as a call of caution, enthusiasm should be limited for publishing effects that are not physiologically relevant.

Topics: Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Coumarins; Estrogen Antagonists; Estrogens; Female; Humans; Polyphenols; Resveratrol; Stilbenes

The breast cancer resistance protein (BCRP/ABCG2) is a drug efflux transporter that can affect the pharmacological and toxicological properties of many molecules. Urolithins, metabolites produced by the gut microbiota from ellagic acid (EA) and ellagitannins, have been acknowledged with in vivo anti-inflammatory and cancer chemopreventive properties. This study evaluated whether urolithins (Uro-A, -B, -C, and -D) and their main phase II metabolites Uro-A sulfate, Uro-A glucuronide, and Uro-B glucuronide as well as their precursor EA were substrates for ABCG2/BCRP. Parental and Bcrp1-transduced MDCKII cells were used for active transport assays. Uro-A and, to a lesser extent, Uro-A sulfate showed a significant increase in apically directed translocation in Bcrp1-transduced cells. Bcrp1 did not show affinity for the rest of the tested compounds. Data were confirmed for murine, human, bovine, and ovine BCRP-transduced subclones as well as with the use of the selective BCRP inhibitor Ko143. The transport inhibition by Uro-A was analyzed by flow cytometry compared to Ko143 using the antineoplastic agent mitoxantrone as a model substrate. Results showed that Uro-A was able to inhibit mitoxantrone transport in a dose-dependent manner. This study reports for the first time that Uro-A and its sulfate conjugate are ABCG2/BCRP substrates. The results suggest that physiologically relevant concentrations of these gut microbiota-derived metabolites could modulate ABCG2/BCRP-mediated transport processes and mechanisms of cancer drug resistance. Further in vivo investigations are warranted.

Topics: Adenosine; Animals; Anti-Inflammatory Agents; Antineoplastic Agents; ATP Binding Cassette Transporter, Subfamily G, Member 2; ATP-Binding Cassette Transporters; Breast Neoplasms; Cattle; Cell Line, Tumor; Coumarins; Diketopiperazines; Dogs; Ellagic Acid; Female; Glucuronides; Heterocyclic Compounds, 4 or More Rings; Humans; Hydrolyzable Tannins; Madin Darby Canine Kidney Cells; Mice; Microbiota; Mitoxantrone; Neoplasm Proteins; Sheep; Sulfates

Urolithins A and B (hydroxy-6H-dibenzo[b,d]pyran-6-one derivatives) are colonic microflora metabolites recently proposed as biomarkers of human exposure to dietary ellagic acid derivatives. Molecular models suggest that urolithins could display estrogenic and/or antiestrogenic activity. To this purpose, both urolithins and other known phytoestrogens (genistein, daidzein, resveratrol, and enterolactone) were assayed to evaluate the capacity to induce cell proliferation on the estrogen-sensitive human breast cancer MCF-7 cells as well as the ability to bind to alpha- and beta-estrogen receptors. Both urolithins A and B showed estrogenic activity in a dose-dependent manner even at high concentrations (40 microM), without antiproliferative or toxic effects, whereas the other phytoestrogens inhibited cell proliferation at high concentrations. Overall, urolithins showed weaker estrogenic activity than the other phytoestrogens. However, both urolithins displayed slightly higher antiestrogenic activity (antagonized the growth promotion effect of 17-beta-estradiol in a dose-dependent manner) than the other phytoestrogens. The IC(50) values for the ERalpha and ERbeta binding assays were 0.4 and 0.75 microM for urolithin A; 20 and 11 microM for urolithin B; 3 and 0.02 for genistein; and 2.3 and 1 for daidzein, respectively; no binding was detected for resveratrol and enterolactone. Urolithins A and B entered into MCF-7 cells and were metabolized to yield mainly urolithin-sulfate derivatives. These results, together with previous studies regarding absorption and metabolism of dietary ellagitannins and ellagic acid in humans, suggest that the gut microflora metabolites urolithins are potential endocrine-disrupting molecules, which could resemble other described "enterophytoestrogens" (microflora-derived metabolites with estrogenic/antiestrogenic activity). Further research is warranted to evaluate the possible role of ellagitannins and ellagic acid as dietary "pro-phytoestrogens".

Topics: Breast Neoplasms; Cell Line, Tumor; Colon; Coumarins; Dose-Response Relationship, Drug; Estrogen Antagonists; Estrogen Receptor alpha; Estrogen Receptor beta; Estrogens; Genistein; Humans; Isoflavones