Nutr Res Pract.  2024 Jun;18(3):436-445. 10.4162/nrp.2024.18.3.436.

A combination of myokines and genistein suppresses cancer stemness in MCF-7 human breast cancer cells

• Kwon H
• Kim Y
• Kim JH

• KMID: 2558515
• DOI: http://doi.org/10.4162/nrp.2024.18.3.436

Abstract

BACKGROUND/OBJECTIVES
Breast cancer is considered a serious health issue worldwide and is influenced by risk factors, including physical inactivity and unhealthy diet. Myokines secreted by muscles during physical activity play a crucial role in cancer development and the immune system. Genistein (Gen), an isoflavone primarily in legumes, induces anti-cancer activity by regulating cancer stem cells (CSCs). Therefore, this study investigated the potential anti-cancer effect of a combination of myokine and Gen on the human breast cancer MCF-7 cells.
MATERIALS/METHODS
MCF-7, a human breast cancer cell line, was used for in vitro study. The cell viability of MCF-7 cells was evaluated in response to treatment with myokines, irisin (Iri), oncostatin M (OSM), and Gen using the MTT assay. Clonogenic and sphere formation assays were used to evaluate the self-renewal capacity of breast CSCs. The mRNA expression levels of stem cell markers were analyzed in MCF-7 breast cancer cells.
RESULTS
Administering Iri or OSM with Gen significantly inhibited the self-renewal capacity of MCF-7 cells. In addition, mRNA expression of breast CSC markers SOX2 and OCT4, which are characteristic of CSCs, was suppressed by both myokine and Gen. However, combining Iri or OSM with Gen was the most effective treatment.
CONCLUSION
These results suggested that combining Iri or OSM with Gen has an additive effect on breast CSCs by regulating self-renewal capacity and expression of CSCs markers. Therefore, the combination of myokines and Gen may have the therapeutic potential for treating breast cancer and improving the quality of life of cancer patients.

Keyword

Breast neoplasms; cancer stem cells; genistein; myokines

Figure

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  Fig. 1 Effects of myokines, Gen, or their combination on MCF-7 cell viability. MCF-7 cells were applied with various concentrations of Iri (0, 10, 20, 40, and 80 nM), OSM (0, 5, 10, 20, and 40 ng/mL), and Gen (0, 100, 125, 150, 175, and 200 μM) for 72 h. (A) Iri, (B) OSM, (C) Gen, and (D) IC50 values of Gen in MCF-7 cells are shown. All data are represented as mean ± SE of the mean from at least 3 independent experiments (n = 3) and analyzed by one-way analysis of variance with Newman-Keuls post hoc test. Different alphabet letters indicate significant differences (P < 0.05).O.D., optical density; Ctrl, control; Iri, irisin; OSM, oncostatin M; Gen, genistein; IC50, half maximal inhibitory concentration.

• 

  Fig. 2 Effects of myokines, Gen, or their combination on colony formation ability of MCF-7 cells. (A) MCF-7 cells were seeded into 6-well plates and treated with Iri, OSM, and Gen. After 10−14 days, colonies were stained and photographed. (B) The number of colonies was counted. All data are represented as mean ± SE of the mean from at least 3 independent experiments (n = 3) and analyzed by one-way analysis of variance with Newman-Keuls post hoc test. Different alphabet letters indicate significant differences (P < 0.05). Data are presented as below: MCF-7 (Iri): Ctrl, control; Iri, 20 nM Iri; OSM, 10 ng/mL OSM; Gen, 117.5 μM Gen; Com, 20 nM Iri + 117.5 μM Gen. MCF-7 (Osm): Ctrl, control; OSM, 10 ng/mL OSM; Gen, 117.5 μM Gen; Com, 10 ng/mL OSM + 117.5 μM Gen.Iri, irisin; Ctrl, control; Gen, genistein; Com, combination; OSM, oncostatin M.

• 

  Fig. 3 Effects of myokines, Gen, or their combination on sphere formation by MCF-7 cells. MCF-7 cells were cultured in poly-HEMA coated 6-well plates and treated with Iri, OSM, Gen, or their combination. Spheres were formed and maintained in sphere media for 10−14 days. (A) Spheres were photographed using phase contrast microscopy (magnification, 100 ×). (B) Several spheres were counted. All data are represented as mean ± SE of the mean from at least 3 independent experiments (n = 3) and analyzed by one-way analysis of variance with Newman-Keuls post hoc test. Different alphabet letters indicate significant difference (P < 0.05).Data are presented as below: MCF-7 (Iri): Ctrl, control; Iri, 20 nM Iri; OSM, 10 ng/mL OSM; Gen, 117.5 μM Gen; Com, 20 nM Iri + 117.5 μM Gen. MCF-7 (Osm): Ctrl, control; OSM, 10 ng/mL OSM; Gen, 117.5 μM Gen; Com, 10 ng/mL OSM + 117.5 μM Gen.Iri, irisin; Ctrl, control; Gen, genistein; Com, combination; OSM, oncostatin M.

• 

  Fig. 4 Effects of myokines, Gen, or their combination on the mRNA expression of CSC markers in MCF-7 cells. MCF-7 cells were treated with myokines (Iri and OSM), Gen, or their combination for 72 h. (A) After treatment of MCF-7 cell lines with (A) Iri and Gen and (B) OSM and Gen, the mRNA expression of SOX2 and OCT4 are presented by quantitative real-time polymerase chain reaction. GAPDH was used as a loading control. All data are represented as mean ± SE of the mean from at least 3 independent experiments (n = 3) and analyzed by 1-way analysis of variance with Newman-Keuls post hoc test. Different alphabet letters indicate significant difference (P < 0.05).Data are presented as below: MCF-7 (Iri): Ctrl, control; Iri, 20 nM Iri; OSM, 10 ng/mL OSM; Gen, 117.5 μM Gen; Com, 20 nM Iri + 117.5 μM Gen. MCF-7 (Osm): Ctrl, control; OSM, 10 ng/mL OSM; Gen, 117.5 μM Gen; Com, 10 ng/mL OSM + 117.5 μM Gen.Ctrl, control; Iri, irisin; Gen, genistein; Com, combination; OSM, oncostatin M.

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