Fresh organically grown pomegranates (Punica granatum L.) of the Wonderful cultivar were processed into three components: fermented juice, aqueous pericarp extract and cold-pressed or supercritical CO2-extracted seed oil. Exposure to additional solvents yielded polyphenol-rich fractions ('polyphenols') from each of the three components. Their actions, and of the crude whole oil and crude fermented and unfermented juice concentrate, were assessed in vitro for possible chemopreventive or adjuvant therapeutic potential in human breast cancer. The ability to effect a blockade of endogenous active estrogen biosynthesis was shown by polyphenols from fermented juice, pericarp, and oil, which inhibited aromatase activity by 60-80%. Fermented juice and pericarp polyphenols, and whole seed oil, inhibited 17-β-hydroxysteroid dehydrogenase Type 1 from 34 to 79%, at concentrations ranging from 100 to 1,000 μg/ml according to seed oil ≫ fermented juice polyphenols > pericarp polyphenols. In a yeast estrogen screen (YES) lyophilized fresh pomegranate juice effected a 55% inhibition of the estrogenic activity of 17-β-estradiol; whereas the lyophilized juice by itself displayed only minimal estrogenic action. Inhibition of cell lines by fermented juice and pericarp polyphenols was according to estrogen-dependent (MCF-7) ≫ estrogen-independent (MB-MDA-231) > normal human breast epithelial cells (MCF-10A). In both MCF-7 and MB-MDA-231 cells, fermented pomegranate juice polyphenols consistently showed about twice the anti-proliferative effect as fresh pomegranate juice polyphenols. Pomegranate seed oil effected 90% inhibition of proliferation of MCF-7 at 100 μg/ml medium, 75% inhibition of invasion of MCF-7 across a Matrigel membrane at 10 μg/ml, and 54% apoptosis in MDA-MB-435 estrogen receptor negative metastatic human breast cancer cells at 50 μg/ml. In a murine mammary gland organ culture, fermented juice polyphenols effected 47% inhibition of cancerous lesion formation induced by the carcinogen 7,12-dimethylbenz[a]anthracene (DMBA). The findings suggest that clinical trials to further assess chemopreventive and adjuvant therapeutic applications of pomegranate in human breast cancer may be warranted.
Bibliographical noteFunding Information:
This work was supported in part through the generous assistance of the Office of the Chief Scientist, State of Israel. N.D. Kim thanks the grant (#HMP-98-D-4-0036) of the Good Health R&D Project, Ministry of Health & Welfare, R.O.K. Thanks also to Professor John McLachlan of the Center for Bioenvironmental Research, Tulane University, for supplying genetically modified yeast for a yeast estrogen screen, and to Dr Janet Price of the Anderson Cancer Center, Texas, USA for a gift of MDA-MB-435 human estrogen receptor negative metastatic breast cancer cells. We are indebted to Jeffrey Marks, master winemaker of Amiad Wineries, Israel, for fermentation of the pomegranate juice, to Agnes Lardet and Phillipe Mengal of Hitex, Vannes, France, for expert supercritical CO2 extraction of the pomegranate seed oil, and to Eli Merom of Kibbutz Sde Eliahu, Israel, many thanks for kindly providing generous amounts of fresh, organically grown pomegranates. We would also like to thank Professor David Gewirtz and Dr Mona Gupta of Virginia Commonwealth University for various support and advice in the preparation of the manuscript. For continuing updates on research relating to pomegranates and cancer, the reader is referred to http://www.rimonest.com.
- 17-β-hydroxysteroid dehydrogenase
- Breast cancer
- Punicic acid
ASJC Scopus subject areas
- Cancer Research