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Breast Cancer Research and Treatment

, Volume 136, Issue 1, pp 21–34 | Cite as

Cytotoxicity of pomegranate polyphenolics in breast cancer cells in vitro and vivo: potential role of miRNA-27a and miRNA-155 in cell survival and inflammation

  • Nivedita Banerjee
  • Stephen Talcott
  • Stephen Safe
  • Susanne U. Mertens-TalcottEmail author
Preclinical Study

Abstract

Several studies have demonstrated that polyphenolics from pomegranate (Punica granatum L.) are potent inhibitors of cancer cell proliferation and induce apoptosis, cell cycle arrest, and also decrease inflammation in vitro and vivo. There is growing evidence that botanicals exert their cytotoxic and anti-inflammatory activities, at least in part, by decreasing specificity protein (Sp) transcription factors. These are overexpressed in breast tumors and regulate genes important for cancer cell survival and inflammation such as the p65 unit of NF-κB. Moreover, previous studies have shown that Pg extracts decrease inflammation in lung cancer cell lines by inhibiting phosphatidylinositol-3,4,5-trisphosphate (PI3K)-dependent phosphorylation of AKT in vitro and inhibiting the activation of NF-kB in vivo. The objective of this study was to investigate the roles of miR-27a–ZBTB10–Sp and miR-155–SHIP-1–PI3K on the anti-inflammatory and cytotoxic activity of pomegranate extract. Pg extract (2.5-50 μg/ml) inhibited growth of BT-474 and MDA-MB-231 cells but not the non-cancer MCF-10F and MCF-12F cells. Pg extract significantly decreased Sp1, Sp3, and Sp4 as well as miR-27a in BT474 and MDA-MB-231 cells and increased expression of the transcriptional repressor ZBTB10. A significant decrease in Sp proteins and Sp-regulated genes was also observed. Pg extract also induced SHIP-1 expression and this was accompanied by downregulation of miRNA-155 and inhibition of PI3K-dependent phosphorylation of AKT. Similar results were observed in tumors from nude mice bearing BT474 cells as xenografts and treated with Pg extract. The effects of antagomirs and knockdown of SHIP-1 by RNA interference confirmed that the anti-inflammatory and cytotoxic effects of Pg extract were partly due to the disruption of both miR-27a–ZBTB10 and miR-155–SHIP-1. In summary, the anticancer activities of Pg extract in breast cancer cells were due in part to targeting microRNAs155 and 27a. Both pathways play an important role in the proliferative/inflammatory phenotype exhibited by these cell lines.

Keywords

Breast cancer Polyphenolics Pomegranate Xenografts Inflammation Cytotoxicity 

Notes

Acknowledgments

We would like to thank Dr. Weston Porter Department Veterinary Integrated Bioscience, at Texas A&M University, College Station, and Texas for providing imaging equipment. Lastly we would like to thank Stefan Wypyszyk at Stiebs LLC (Kirkland, WA) for kindly supplying the pomegranate juice. Financial support for this research has been provided by the National Institutes of Health (KOIATOO 4597 to SMT).

Conflict of interest

The authors have no conflicts of interest to declare.

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Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Nivedita Banerjee
    • 1
    • 2
  • Stephen Talcott
    • 2
  • Stephen Safe
    • 1
    • 3
  • Susanne U. Mertens-Talcott
    • 1
    • 2
    • 3
    • 4
    Email author
  1. 1.Interdisciplinary Program of ToxicologyTexas A&M UniversityCollege StationUSA
  2. 2.Department of Nutrition & Food ScienceTexas A&M UniversityCollege StationUSA
  3. 3.Department of Veterinary Physiology & Pharmacology, Texas A&M UniversityCollege of Veterinary MedicineCollege StationUSA
  4. 4.Institute for Obesity Research and Program Evaluation, Texas A&M UniversityCollege StationUSA

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