Evaluation of Manuka Honey Estrogen Activity Using the MCF-7 Cell Proliferation Assay

Katheleen Henderson, Tahrir Aldhirgham, Poonam Singh - Nee Nigam, Richard K. Owusu-Apenten

Research output: Contribution to journalArticle

Abstract

Aims: To assess the estrogenic activity of Manuka honey using the MCF7 cell proliferation assay.Study Design: In-vitro cell based E-screen.Place and Duration of Study: Ulster University, Coleraine, UK, September 2015 to September 2016.Methodology: Manuka honey (UMF15+) was characterized for total phenolic content (TPC) using the Folin-Ciocalteu assay and antioxidant power, using the 2, 2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assay. Estrogenic activity was assessed using MCF-7 cells cultured in DMEM-F2 phenol red-free media supplemented with 10% charcoal stripped FBS and evaluated using Sulforhodamine B (SRB) colorimetric assay. All experiments were conducted in triplicate (n-12-48) and genistein was the positive control. The effect of Manuka honey (UMF15+) treatment on intracellular reactive oxygen species (ROS) was measured using the 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA) assay. Results: Manuka honey (UMF15+) antioxidant power was related to total phenols content. MCF7 growth promotion occurred at very low concentrations of honey (5x10-6-5x10-3% v/v honey; or 2.75 x10-10M - 2.75 x10-7 M TPC) indicative of estrogenic activity whilst higher concentrations of honey (>0.5% v/v) were inhibitory. Similarly, the genistein positive control demonstrated estrogenic activity indicated by MCF-7 cell growth at low concentrations (5x10-9-5x10-8 M) and toxicity at high concentrations. Estrogenic characteristics were quantified in terms of the relative proliferative potency (RPP) and relative proliferative effect (RPE) for Manuka honey of 18% and 22.5-27.5%, respectively. For genistein RPP was 0.1% and RPE was 70% compared to values of 100% for estradiol. Intracellular ROS increased for MCF-7 cells treated with increasing honey concentrations. Conclusion: Manuka honey (UMF15+) exhibits estrogenic activity monitored as growth promotion of MCF-7 breast cancer cells with estrogenic parameters being comparable to values reported for some purified flavonoids. Treatment of MCF-7 cells produced a dose-dependent rise in intracellular ROS.
LanguageEnglish
Pages1-11
JournalJournal of Advances in Biology and Biotechnology
Volume10
Issue number3
Early online date1 Dec 2016
DOIs
Publication statusE-pub ahead of print - 1 Dec 2016

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Honey
MCF-7 Cells
Estrogens
Cell Proliferation
Genistein
Reactive Oxygen Species
lissamine rhodamine B
Growth
Antioxidants
Phenolsulfonphthalein
Sulfonic Acids
Phenols
Charcoal
Flavonoids
Estradiol
Breast Neoplasms

Keywords

  • Manuka honey
  • estrogenic activity
  • breast cancer
  • antioxidant activity
  • E-SCREEN
  • MCF-7.

Cite this

Henderson, Katheleen ; Aldhirgham, Tahrir ; Singh - Nee Nigam, Poonam ; Owusu-Apenten, Richard K. / Evaluation of Manuka Honey Estrogen Activity Using the MCF-7 Cell Proliferation Assay. In: Journal of Advances in Biology and Biotechnology. 2016 ; Vol. 10, No. 3. pp. 1-11.
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abstract = "Aims: To assess the estrogenic activity of Manuka honey using the MCF7 cell proliferation assay.Study Design: In-vitro cell based E-screen.Place and Duration of Study: Ulster University, Coleraine, UK, September 2015 to September 2016.Methodology: Manuka honey (UMF15+) was characterized for total phenolic content (TPC) using the Folin-Ciocalteu assay and antioxidant power, using the 2, 2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assay. Estrogenic activity was assessed using MCF-7 cells cultured in DMEM-F2 phenol red-free media supplemented with 10{\%} charcoal stripped FBS and evaluated using Sulforhodamine B (SRB) colorimetric assay. All experiments were conducted in triplicate (n-12-48) and genistein was the positive control. The effect of Manuka honey (UMF15+) treatment on intracellular reactive oxygen species (ROS) was measured using the 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA) assay. Results: Manuka honey (UMF15+) antioxidant power was related to total phenols content. MCF7 growth promotion occurred at very low concentrations of honey (5x10-6-5x10-3{\%} v/v honey; or 2.75 x10-10M - 2.75 x10-7 M TPC) indicative of estrogenic activity whilst higher concentrations of honey (>0.5{\%} v/v) were inhibitory. Similarly, the genistein positive control demonstrated estrogenic activity indicated by MCF-7 cell growth at low concentrations (5x10-9-5x10-8 M) and toxicity at high concentrations. Estrogenic characteristics were quantified in terms of the relative proliferative potency (RPP) and relative proliferative effect (RPE) for Manuka honey of 18{\%} and 22.5-27.5{\%}, respectively. For genistein RPP was 0.1{\%} and RPE was 70{\%} compared to values of 100{\%} for estradiol. Intracellular ROS increased for MCF-7 cells treated with increasing honey concentrations. Conclusion: Manuka honey (UMF15+) exhibits estrogenic activity monitored as growth promotion of MCF-7 breast cancer cells with estrogenic parameters being comparable to values reported for some purified flavonoids. Treatment of MCF-7 cells produced a dose-dependent rise in intracellular ROS.",
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Evaluation of Manuka Honey Estrogen Activity Using the MCF-7 Cell Proliferation Assay. / Henderson, Katheleen; Aldhirgham, Tahrir; Singh - Nee Nigam, Poonam; Owusu-Apenten, Richard K.

In: Journal of Advances in Biology and Biotechnology, Vol. 10, No. 3, 01.12.2016, p. 1-11.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Evaluation of Manuka Honey Estrogen Activity Using the MCF-7 Cell Proliferation Assay

AU - Henderson, Katheleen

AU - Aldhirgham, Tahrir

AU - Singh - Nee Nigam, Poonam

AU - Owusu-Apenten, Richard K.

PY - 2016/12/1

Y1 - 2016/12/1

N2 - Aims: To assess the estrogenic activity of Manuka honey using the MCF7 cell proliferation assay.Study Design: In-vitro cell based E-screen.Place and Duration of Study: Ulster University, Coleraine, UK, September 2015 to September 2016.Methodology: Manuka honey (UMF15+) was characterized for total phenolic content (TPC) using the Folin-Ciocalteu assay and antioxidant power, using the 2, 2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assay. Estrogenic activity was assessed using MCF-7 cells cultured in DMEM-F2 phenol red-free media supplemented with 10% charcoal stripped FBS and evaluated using Sulforhodamine B (SRB) colorimetric assay. All experiments were conducted in triplicate (n-12-48) and genistein was the positive control. The effect of Manuka honey (UMF15+) treatment on intracellular reactive oxygen species (ROS) was measured using the 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA) assay. Results: Manuka honey (UMF15+) antioxidant power was related to total phenols content. MCF7 growth promotion occurred at very low concentrations of honey (5x10-6-5x10-3% v/v honey; or 2.75 x10-10M - 2.75 x10-7 M TPC) indicative of estrogenic activity whilst higher concentrations of honey (>0.5% v/v) were inhibitory. Similarly, the genistein positive control demonstrated estrogenic activity indicated by MCF-7 cell growth at low concentrations (5x10-9-5x10-8 M) and toxicity at high concentrations. Estrogenic characteristics were quantified in terms of the relative proliferative potency (RPP) and relative proliferative effect (RPE) for Manuka honey of 18% and 22.5-27.5%, respectively. For genistein RPP was 0.1% and RPE was 70% compared to values of 100% for estradiol. Intracellular ROS increased for MCF-7 cells treated with increasing honey concentrations. Conclusion: Manuka honey (UMF15+) exhibits estrogenic activity monitored as growth promotion of MCF-7 breast cancer cells with estrogenic parameters being comparable to values reported for some purified flavonoids. Treatment of MCF-7 cells produced a dose-dependent rise in intracellular ROS.

AB - Aims: To assess the estrogenic activity of Manuka honey using the MCF7 cell proliferation assay.Study Design: In-vitro cell based E-screen.Place and Duration of Study: Ulster University, Coleraine, UK, September 2015 to September 2016.Methodology: Manuka honey (UMF15+) was characterized for total phenolic content (TPC) using the Folin-Ciocalteu assay and antioxidant power, using the 2, 2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assay. Estrogenic activity was assessed using MCF-7 cells cultured in DMEM-F2 phenol red-free media supplemented with 10% charcoal stripped FBS and evaluated using Sulforhodamine B (SRB) colorimetric assay. All experiments were conducted in triplicate (n-12-48) and genistein was the positive control. The effect of Manuka honey (UMF15+) treatment on intracellular reactive oxygen species (ROS) was measured using the 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA) assay. Results: Manuka honey (UMF15+) antioxidant power was related to total phenols content. MCF7 growth promotion occurred at very low concentrations of honey (5x10-6-5x10-3% v/v honey; or 2.75 x10-10M - 2.75 x10-7 M TPC) indicative of estrogenic activity whilst higher concentrations of honey (>0.5% v/v) were inhibitory. Similarly, the genistein positive control demonstrated estrogenic activity indicated by MCF-7 cell growth at low concentrations (5x10-9-5x10-8 M) and toxicity at high concentrations. Estrogenic characteristics were quantified in terms of the relative proliferative potency (RPP) and relative proliferative effect (RPE) for Manuka honey of 18% and 22.5-27.5%, respectively. For genistein RPP was 0.1% and RPE was 70% compared to values of 100% for estradiol. Intracellular ROS increased for MCF-7 cells treated with increasing honey concentrations. Conclusion: Manuka honey (UMF15+) exhibits estrogenic activity monitored as growth promotion of MCF-7 breast cancer cells with estrogenic parameters being comparable to values reported for some purified flavonoids. Treatment of MCF-7 cells produced a dose-dependent rise in intracellular ROS.

KW - Manuka honey

KW - estrogenic activity

KW - breast cancer

KW - antioxidant activity

KW - E-SCREEN

KW - MCF-7.

U2 - 10.9734/JABB/2016/29887

DO - 10.9734/JABB/2016/29887

M3 - Article

VL - 10

SP - 1

EP - 11

JO - Journal of Advances in Biology and Biotechnology

T2 - Journal of Advances in Biology and Biotechnology

JF - Journal of Advances in Biology and Biotechnology

SN - 2394-1081

IS - 3

ER -