Manuka Honey Inhibits Human Breast Cancer Progression in Preclinical Models
<p>Manuka honey reduces the proliferation of ER-positive human breast cancer cells in vitro. ER-positive MCF-7 breast cancer cells and TNBC MDA-MB-231 cells were incubated in the presence of increasing concentrations of either (<b>A</b>) Manuka honey at 0.0 to 5.0% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) or (<b>B</b>) Manuka powder at 0.0–16%. After 72 h, cell counts were performed using an MTS assay and by manual cell counts. The figures show tumor cell proliferation expressed as the mean percentage of vehicle-treated controls with SEM. Experiments were performed at least three times in independent experiments. (<b>C</b>) Manuka honey reduces the proliferation of MCF-7 cancer cells but not that of non-malignant mammary cells in vitro and enhances the antitumor action of the antiestrogen tamoxifen. Human MCF-7 tumor cells and non-malignant HMECs were cultured in vitro with 2.5% <span class="html-italic">w/v</span> Manuka honey (MH), 10 μM tamoxifen (TM), or both agents combined for 48 h. Cell proliferation was then quantitated and expressed as a percentage of that recorded in appropriate vehicle-treated controls. A higher 5% <span class="html-italic">w/v</span> MH concentration was also tested without a significant effect on HMEC proliferation. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.01, n > 3.</p> "> Figure 2
<p>Induction of apoptosis of breast cancer cells by Manuka honey. (<b>A</b>) MCF-7 cells were treated with vehicle control (CON), 2.5% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) (MH 2.5) or 5% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) (MH 5.0) Manuka honey, camptothecin 1 μM (Cam), 5% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) (Dex) dextrose, or 5% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) (HMesq) Mesquite honey. After 24 and/or 48 h, cells were harvested and stained with Annexin V and 7-AAD to assess early and late apoptosis. Treatments with Manuka honey, particularly at 5.0% (<span class="html-italic">w</span>/<span class="html-italic">v</span>), elicited significant increments in apoptotic cells as compared to controls (* <span class="html-italic">p</span> < 0.05). Camptothecin, a positive control drug, elicited a similar increase in late apoptotic cells after 48 h, while treatment of MCF-7 cells with dextrose or Mesquite honey did not exhibit comparable increments in the numbers of apoptotic cells. (<b>B</b>) Treatment of MCF-7 cells with Manuka honey elicits increased poly (ADP-ribose) polymerase (PARP) cleavage. Cells were treated in vitro for 48 h with either control vehicle or Manuka honey at 0.6, 2.5, or 5.0% (<span class="html-italic">w</span>/<span class="html-italic">v</span>). Camptothecin was also used as a positive control.</p> "> Figure 3
<p>Manuka honey activates AMPK signaling and inhibits mTOR and STAT3 downstream signaling. (<b>A</b>) MCF7 cells were treated with increasing concentrations of Manuka honey (0–5%) and 5% Mesquite honey as control. After 24 h, cells were lysed and immunoblotted with specific antibodies. (<b>B</b>) MCF7 cells were treated with increasing concentrations of Manuka honey 0.3–5% (<span class="html-italic">w</span>/<span class="html-italic">v</span>). After 24 h, cells were lysed, and whole cell extracts were resolved by PAGE and immunoblotted with specific antibodies.</p> "> Figure 4
<p>Antitumor activity of Manuka honey in human breast cancer xenografts in vivo. Ovariectomized nude mice (nu<sup>−</sup>/nu<sup>−</sup>, Charles Rivers) with estradiol supplements were implanted with MCF-7 tumor xenografts SQ and treated with Manuka honey or control administered by oral gavage after tumors achieved a size of 50–75 cm<sup>3</sup>. Oral gavage (0.1 mL volume) with 50% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) Manuka honey or control was performed twice daily from days 1 to 14, then once daily thereafter to day 42. Treatment with Manuka honey administered orally elicited a significant suppression of MCF-7 xenograft progression as compared to controls (** <span class="html-italic">p</span> < 0.01) n = 5–7 mice per group.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents
2.2. Cell Lines
2.3. Cell Proliferation Assay
2.4. Western Blot Analysis
2.5. Annexin V Staining Assay
2.6. Human Tumor Xenografts in Nude Mice
2.7. Statistics
3. Results
3.1. Manuka Honey Reduces the Proliferation of Human MCF-7 Breast Cancer Cells In Vitro
3.2. MH Induces Apoptosis in MCF-7 Breast Cancer Cells In Vitro
3.3. Manuka Honey Induces PARP-Cleavage Leading to Tumor Cell Apoptosis
3.4. Manuka Honey Activates AMPK and Inhibits Downstream PI3K/AKT/mTOR Signaling
3.5. Manuka Honey Inhibits STAT-3 Phosphorylation
3.6. Antitumor Activity of Manuka Honey in Human Breast Cancer Xenografts In Vivo
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Márquez-Garbán, D.C.; Yanes, C.D.; Llarena, G.; Elashoff, D.; Hamilton, N.; Hardy, M.; Wadehra, M.; McCloskey, S.A.; Pietras, R.J. Manuka Honey Inhibits Human Breast Cancer Progression in Preclinical Models. Nutrients 2024, 16, 2369. https://doi.org/10.3390/nu16142369
Márquez-Garbán DC, Yanes CD, Llarena G, Elashoff D, Hamilton N, Hardy M, Wadehra M, McCloskey SA, Pietras RJ. Manuka Honey Inhibits Human Breast Cancer Progression in Preclinical Models. Nutrients. 2024; 16(14):2369. https://doi.org/10.3390/nu16142369
Chicago/Turabian StyleMárquez-Garbán, Diana C., Cristian D. Yanes, Gabriela Llarena, David Elashoff, Nalo Hamilton, Mary Hardy, Madhuri Wadehra, Susan A. McCloskey, and Richard J. Pietras. 2024. "Manuka Honey Inhibits Human Breast Cancer Progression in Preclinical Models" Nutrients 16, no. 14: 2369. https://doi.org/10.3390/nu16142369