Agarol, an ergosterol derivative from Agaricus blazei, induces caspase-independent apoptosis in human cancer cells
Agarol, an ergosterol derivative from Agaricus blazei, induces caspase-independent apoptosis in human cancer cells
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DOI：
10.3892/ijo.2016.3391
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发表时间：
2016-04-01
影响因子：
5.2
通讯作者：
Hidemi, Rikiishi
中科院分区：
文献类型：
--
作者：
Shimizu, Takamitsu;Kawai, Junya;Hidemi, Rikiishi
巴西蘑菇Agaricus blazei（A. blazei）是一种具有多种生物作用和活性成分的蘑菇。我们从A. blazei，麦角甾醇衍生物，并将其命名为“Agarol”。使用经琼脂醇处理的A549、MKN 45、HSC-3和HSC-4人癌细胞系，通过MTT测定法测定琼脂醇的细胞毒性作用。流式细胞仪检测细胞凋亡。活性氧（ROS）水平和线粒体膜电位（Δ Psim）也通过流式细胞术测定。Western blot分析用于定量糖尿病相关蛋白的表达。与其他p53突变细胞系（HSC-3和HSC-4）相比，琼脂醇主要诱导两种p53野生型细胞系（A549和MKN 45）的凋亡。进一步的机制研究表明，细胞凋亡的诱导与ROS产生增加、Δ Psim降低、凋亡诱导因子（AIF）从线粒体释放到胞质溶胶、Bax上调和Bcl-2下调相关。Caspase-3活性没有增加，caspase抑制剂z-VAD-falanine不能抑制Agarol诱导的细胞凋亡。这些发现表明，琼脂醇诱导caspase非依赖性凋亡的人癌细胞通过线粒体途径。在异种移植小鼠模型中证实了Agarol的体内抗癌活性。本研究提示了Agarol诱导人癌细胞凋亡的分子机制，并表明Agarol作为抗癌剂的潜在用途。
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 (Delta Psi m) 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 Delta Psi m, 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.