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Cell Biochemistry and Biophysics

, Volume 63, Issue 1, pp 85–96 | Cite as

Steroidal Metabolites Transformed by Marchantia polymorpha Cultures Block Breast Cancer Estrogen Biosynthesis

  • Mohamed-Elamir F. HegazyEmail author
  • Amira M. Gamal-Eldeen
  • Ali M. El-Halawany
  • Abou El-Hamd H. Mohamed
  • Paul W. Paré
Original Paper

Abstract

Suspension of cultured cells of Marchantia polymorpha have the potential to hydrogenate the olefinic bonds present in androst-1,4-dien-3,17-dione (boldione, 1) to afford dihydroandrost-3,17-dione derivatives including: androst-4-ene-3,17-dione (androstenedione, 4-AD, 2), 5α-androstane-3,17-dione (androstenedione, AD, 4), and the less abundant metabolite 5α-androst-1-ene-3,17-dione (1-androstenedione, 1-AD, 3). After isolation and purification, these metabolites were characterized on the basis of spectroscopic analyses using 1D and 2D NMR as well as mass spectrometry. Cytotoxicity of the biotransformation products against breast adenocarcinoma cells (MCF-7) was assessed by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay and cell death (apoptosis or necrosis) was assayed by acridine orange/ethidium bromide staining. Aromatase (cytochrome P450 19 enzyme, CYP19) inhibitory activity was measured by a tritiated water release assay and by direct measurement of bio-transformed steroids using the tritium labeled substrate 3H-androst-4-ene-3,17-dione. CYP19 mRNA expression in MCF-7 cells was analyzed by real-time PCR. Steroidal products 3 and 4 revealed a highly significant inhibition of MCF-7 cell growth that was predominantly due to apoptosis not necrosis. Steroidal products 3 and 4 are both potent inhibitors of aromatase activity and CYP19 mRNA expression, while 2 is a known substrate for aromatase. These data establish that metabolites 3 and 4 are potent chemical agents against breast cancer via aromatase inhibitory mechanism. Results were interpreted via virtual docking of the biotransformation products to the human placental aromatase active site.

Keywords

Cultured plant cells Hydrogenation Androst-1,4-dien-3,17-dione Anticancer activity Aromatase inhibitory activity 

Notes

Acknowledgments

This study was funded by the National Research Center, Cairo, Egypt.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Mohamed-Elamir F. Hegazy
    • 1
    • 2
    Email author
  • Amira M. Gamal-Eldeen
    • 3
  • Ali M. El-Halawany
    • 4
  • Abou El-Hamd H. Mohamed
    • 5
  • Paul W. Paré
    • 6
  1. 1.Chemistry of Medicinal Plants Department, and Pharmaceutical Laboratory, Center of Excellence for Advanced SciencesNational Research CenterCairoEgypt
  2. 2.Kyoto Pharmaceutical UniversityKyotoJapan
  3. 3.Cancer Biology Laboratory, Center of Excellence for Advanced Sciences, and Biochemistry DepartmentNational Research CenterCairoEgypt
  4. 4.Department of Pharmacognosy, Faculty of PharmacyCairo UniversityCairoEgypt
  5. 5.Department of Chemistry, Aswan-Faculty of ScienceSouth Valley UniversityAswânEgypt
  6. 6.Department of Chemistry and BiochemistryTexas Tech UniversityLubbockUSA

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