Apigenin drives the production of reactive oxygen species and initiates a mitochondrial mediated cell death pathway in prostate epithelial cells
Apigenin drives the production of reactive oxygen species and initiates a mitochondrial mediated cell death pathway in prostate epithelial cells
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DOI：
10.1002/pros.20167
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发表时间：
2005-05-01
期刊：
影响因子：
2.8
通讯作者：
Watson, RWG
中科院分区：
文献类型：
--
作者：
Morrissey, C;O'Neill, A;Watson, RWG
背景植物雌激素可减少前列腺癌的肿瘤发生。我们筛选了5种植物雌激素对PWR-1 E、LNCaP、PC-3和DU 145前列腺上皮细胞体外生长和凋亡的影响。我们使用结晶紫分析、流式细胞术分析和TUNEL法评估细胞数量、增殖和凋亡。我们特别关注芹菜素，评估钙蛋白酶、丝氨酸蛋白酶、半胱天冬酶、雌激素受体和神经酰胺合成酶抑制剂阻断芹菜素诱导的细胞凋亡的能力。我们还分析了caspase 3，7，8，9，Bcl-2，Bax，Bid和细胞色素C的Western分析，和线粒体通透性和活性氧产生的流式细胞术使用mitosensor（TM）和DCFH-DA，分别。芹菜素和水飞蓟宾显着减少细胞数量，芹菜素诱导PWR-1 E，LNCaP，PC-3和DU 145细胞凋亡。PC-3和DU 145细胞对芹菜素诱导的凋亡的敏感性低于LNCaP和PWR-1 E细胞。芹菜素诱导细胞凋亡是半胱天冬酶依赖的。芹菜素产生活性氧，线粒体Bcl-2表达的损失，线粒体通透性，细胞色素C释放，和半胱天冬酶3，7，8，和9的裂解和伴随的细胞凋亡蛋白，cIAP-2的抑制剂的裂解。Bcl-2在LNCaP B10细胞中的过表达降低了芹菜素的凋亡作用。芹菜素通过线粒体介导的细胞死亡途径诱导前列腺上皮细胞死亡。Bcl-2在抑制芹菜素诱导的前列腺上皮细胞死亡中具有作用。(c)2004 Wiley-Liss，Inc.
BACKGROUND. Phytoestrogens may reduce tumorigenesis in prostate cancer. We screened five phytoestrogens for their effect on cell growth and apoptosis in PWR-1E, LNCaP, PC-3, and DU145 prostate epithelial cells in vitro.METHODS. We assessed cell number, proliferation, and apoptosis using crystal violet assays, flow cytometric analysis, and TUNEL. Focusing specifically on apigenin we assessed the ability of calpain, serine protease, caspase, estrogen receptor, and ceramide synthase inhibitors to block apigenin induced apoptosis. We also analyzed caspase 3, 7,8, 9, Bcl-2, Bax, Bid, and cytochrome C by Western analysis, and mitochondrial permeability and reactive oxygen species production by flow cytometry using mitosensor (TM) and DCFH-DA, respectively.RESULTS. Apigenin and silybinin significantly reduced cell number, with apigenin inducing apoptosis in PWR-1E, LNCaP, PC-3, and DU145 cells. The PC-3 and DU145 cells were less susceptible to apigenin induced apoptosis then LNCaP and PWR-1E cells. The induction of apoptosis by apigenin was caspase dependent. Apigenin generated reactive oxygen species, a loss of mitochondrial Bcl-2 expression, mitochondrial permeability, cytochrome C release, and the cleavage of caspase 3, 7, 8, and 9 and the concomitant cleavage of the inhibitor of apoptosis protein, cIAP-2. The overexpression of Bcl-2 in LNCaP B10 cells reduced the apoptotic effects of apigenin.CONCLUSIONS. Apigenin induces cell death in prostate epithelial cells using a mitochondrial mediated cell death pathway. Bcl-2 has a role in inhibiting apigenin induced cell death in prostate epithelial cells. (c) 2004 Wiley-Liss, Inc.