Urolithin A induces prostate cancer cell death in p53-dependent and in p53-independent manner

• Published in: European journal of nutrition Vol. 59; no. 4; pp. 1607 - 1618
• Main Authors: Mohammed Saleem, Yasir I., Albassam, Hussam, Selim, Mustafa
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.06.2020; Springer Nature B.V
• Subjects: Annexin V; Apoptosis; Bioactive compounds; Blotting, Western; Cell Culture Techniques; Cell death; Cell Death - drug effects; Cell viability; Chemistry; Chemistry and Materials Science; Coumarins - pharmacology; Cyclin-dependent kinase inhibitor p21; Flow Cytometry; Gene expression; Genotypes; GTP-binding protein; Humans; Immunoblotting; Immunoprecipitation; Intestinal microflora; Male; MDM2 protein; Microbiota; Nutrition; Original Contribution; p53 Protein; Polymerase Chain Reaction; Polyphenols; Prostate cancer; Prostatic Neoplasms - genetics; Prostatic Neoplasms - metabolism; Protein expression; Proteins; Tumor cell lines; Tumor suppressor genes; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Western blotting; XIAP protein; MDM2; Polyphenols; Urolithin A; Prostate cancer; p53
• ISSN: 1436-6207; 1436-6215
• DOI: 10.1007/s00394-019-02016-2

Purpose Pomegranate and walnuts are widely consumed dietary sources and contain several bioactive compounds, including the ellagitannins (ETs). ETs are polyphenols that are metabolized in the gut microbiota to urolithin A (UA). p53 is a tumor suppressor that lost its activity through MDM2 activation in about half cancers. The purpose of this study was to investigate the influence of UA on the p53-MDM2 interaction pathway in prostate cancer cell lines. Methods Three human prostate cancer cell lines were used that harbor different p53 genotypes; LNCaP (p53 +/+ ), 22RV1(p53 −/+ ) and PC3 (p53 −/− ). Cell viability was determined by CellTiter-Glo Luminescent assay. Apoptosis was confirmed by measuring annexin V by flow cytometry. The expression of p53, its target proteins, and apoptotic markers were measured by western blotting. Real-time qPCR was used to measure the gene expression of p21, a main target gene of p53. Co-immunoprecipitation–immunoblotting was used to assess the inhibition of interactions between p53 and MDM2 and to assess the effect of UA on MDM2-mediated p53 polyubiquitination. Results We found UA inhibited CaP cells’ viability and induced apoptosis. For 22RV1 and LNCaP, we found UA increased p53 protein expression and its main target protein, p21, and MDM2, forming an autoregulatory feedback loop. In addition, UA increased the p53 proapoptotic proteins PUMA and NOXA. Moreover, UA inhibited the interaction between p53 and MDM2 and inhibited MDM2-mediated p53 polyubiquitination. UA downregulated MDM2 and XIAP protein expression in PC3 cells and upregulated p21 and p14ARF in a p53-independent manner. Conclusion The influencing of UA on p53-MDM2 pathway may partly contribute to its anticancer effect.