Agarol, an ergosterol derivative from Agaricus blazei, induces caspase-independent apoptosis in human cancer cells

Shimizu,Takamitsu; Kawai,Junya; Ouchi,Kenji; Kikuchi,Haruhisa; Osima,Yoshiteru; Hidemi,Rikiishi

doi:10.3892/ijo.2016.3391

Agarol, an ergosterol derivative from Agaricus blazei, induces caspase-independent apoptosis in human cancer cells

Authors:
- Takamitsu Shimizu
- Junya Kawai
- Kenji Ouchi
- Haruhisa Kikuchi
- Yoshiteru Osima
- Rikiishi Hidemi
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Affiliations: Mushroom Research Laboratory, Hokuto Corporation, Nagano 381-0008, Japan, Laboratory of Natural Product Chemistry, Tohoku University Graduate School of Pharmaceutical Sciences, Sendai 980-8578, Japan, Department of Microbiology and Immunology, Tohoku University Graduate School of Dentistry, Sendai 980-8575, Japan
Published online on: February 15, 2016 https://doi.org/10.3892/ijo.2016.3391
Pages: 1670-1678

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Abstract

Agaricus blazei (A. blazei) is a mushroom with many biological effects and active ingredients. We purified a tumoricidal substance from A. blazei, an ergosterol derivative, and named it ‘Agarol’. Cytotoxic effects of Agarol were determined by the MTT assay using A549, MKN45, HSC-3, and HSC-4 human carcinoma cell lines treated with Agarol. Apoptosis was detected by flow cytometry analysis. Reactive oxygen species (ROS) levels and mitochondria membrane potential (∆ψm) were also determined by flow cytometry. Western blot analysis was used to quantify the expression of apoptosis-related proteins. Agarol predominantly induced apoptosis in two p53-wild cell lines (A549 and MKN45) compared to the other p53-mutant cell lines (HSC-3 and HSC-4). Further mechanistic studies revealed that induction of apoptosis is associated with increased generation of ROS, reduced ∆ψm, release of apoptosis-inducing factor (AIF) from the mitochondria to the cytosol, upregulation of Bax, and downregulation of Bcl-2. Caspase-3 activities did not increase, and z-VAD-fmk, a caspase inhibitor, did not inhibit the Agarol-induced apoptosis. These findings indicate that Agarol induces caspase-independent apoptosis in human carcinoma cells through a mitochondrial pathway. The in vivo anticancer activity of Agarol was confirmed in a xenograft murine model. This study suggests a molecular mechanism by which Agarol induces apoptosis in human carcinoma cells and indicates the potential use of Agarol as an anticancer agent.

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