Pleurotus highking Mushroom Induces Apoptosis by Altering the Balance of Proapoptotic and Antiapoptotic Genes in Breast Cancer Cells and Inhibits Tumor Sphere Formation.

Md Anwarul Haque,Md Anwar Ul Islam

doi:10.3390/medicina55110716

Md Anwarul Haque, Md Anwar Ul Islam

Open Access

https://doi.org/10.3390/medicina55110716

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Abstract

Background and objectives: Mushrooms that have medicinal properties are part of many traditional diets. The aim of the present study was to use the human breast cancer cell line MCF-7 to investigate the anticancer activity of Pleurotus highking mushroom purified extract fraction-III (PEF-III) and to elucidate the possible mechanism of that activity. Materials and Methods: The effects of PEF-III on cell proliferation and viability were evaluated by a colony formation assay and an MTT assay, respectively. Cell morphological changes, annexin-V phycoerythrin and propidium iodide (PI) staining, DNA fragmentation, and caspase 3/7 activity assays were performed to determine the induction of apoptosis by PEF-III. The genes responsible for regulation of apoptosis were analyzed by means of Western blot analysis. In vitro tumor sphere formation assay was performed using a 3D sphere culture system. Results: PEF-III significantly reduced the proliferation and viability of MCF-7 cells. Cell shrinkage and rounding, and annexin-V phycoerythrin and PI staining followed by flow cytometry indicated that the cell death was due to apoptosis. Additionally, a laddering DNA pattern and increased levels of caspase-3/7 enzyme also corroborated the notion of apoptosis-mediated cell death. This incidence was further confirmed by upregulation of proapoptotic genes (p53 and its target gene, Bax) and downregulation of the expression of an antiapoptotic gene (Bcl-2). PEF-III also reduced the size and number of the tumor spheres in 3D culture conditions. Conclusions: The anticancer activity of PEF-III is due to induction of apoptosis by a shift in the balance of proapoptotic and antiapoptotic genes. Therefore, the findings of the present study may open a path to exploring potential drug candidates from the P. highking mushroom for combating breast cancer.

Highlights

Cancer is one of the most devastating diseases, posing the threat of mortality to individuals worldwide despite promising advances in medical diagnosis and treatment [1]
Annexin-V phycoerythrin and propidium iodide (PI) staining followed by flow cytometry indicated that the cell death was due to apoptosis
The anticancer activity of purified extract fraction-III (PEF-III) is due to induction of apoptosis by a shift in the balance of proapoptotic and antiapoptotic genes

Summary

Introduction

Cancer is one of the most devastating diseases, posing the threat of mortality to individuals worldwide despite promising advances in medical diagnosis and treatment [1]. Considerable attention has been focused on screening anticancer compounds from natural sources, including from medicinal plants [3] As part of such a study, we focused on an edible and medicinal species of the oyster mushroom, the Pleurotus highking. Oyster mushrooms have been considered throughout the world as a functional food and folk medicine [4] They contain diverse biomolecules that are used for the treatment of various diseases including cancer [5]. A laddering DNA pattern and increased levels of caspase-3/7 enzyme corroborated the notion of apoptosis-mediated cell death This incidence was further confirmed by upregulation of proapoptotic genes (p53 and its target gene, Bax) and downregulation of the expression of an antiapoptotic gene (Bcl-2). The findings of the present study may open a path to exploring potential drug candidates from the P. highking mushroom for combating breast cancer

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