Investigation of ouabain-induced anticancer effect in human androgen-independent prostate cancer PC-3 cells
Investigation of ouabain-induced anticancer effect in human androgen-independent prostate cancer PC-3 cells
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DOI：
10.1016/j.bcp.2003.10.013
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发表时间：
2004-02-15
影响因子：
5.8
通讯作者：
Guh, JH
中科院分区：
文献类型：
--
作者：
Huang, YT;Chueh, SC;Guh, JH
为了确定强心苷在雄激素非依赖性前列腺癌中的治疗潜力，我们研究了哇巴因诱导的细胞毒性作用以及PC-3细胞中的信号通路。线粒体MTT法显示哇巴因具有时间和浓度依赖性的细胞毒作用，其有效阈值浓度为纳摩尔。在浓度小于10 nM时，哇巴因诱导线粒体活性降低，直至进行7小时暴露，而早在用高浓度哇巴因处理细胞2小时时，它诱导线粒体功能快速下降，这表明哇巴因作用涉及两种不同的机制。结果表明，低浓度和高浓度的哇巴因均能抑制PC-3细胞Na ~+-K ~+ ATP酶活性，引起Ca ~（45）（2+）内流。高浓度的哇巴因可引起线粒体膜电位（A，）的显著和时间依赖性的损失，活性氧（ROS）的持续产生，以及严重的凋亡反应。哇巴因还诱导Par-4（前列腺凋亡反应4）表达的增加。此外，一个反义的，但不是无义的，寡聚体对Par-4的表达显着抑制低浓度哇巴因诱导的细胞毒性。这表明哇巴因诱导两种模式的细胞毒作用在人前列腺癌PC-3细胞中的非依赖性。低浓度的哇巴因诱导PC-3细胞Par-4表达增加并敏化细胞毒性;而高浓度的哇巴因诱导PC-3细胞中DeltaPsi（m）的损失、持续的ROS产生和严重的凋亡。(C)2003年爱思唯尔公司All rights reserved.
To determine the therapeutic potential of cardiac glycosides in androgen-independent prostate cancer, we examined ouabain-induced cytotoxic effect as well as the signaling pathways in PC-3 cells. Ouabain induced a time- and concentration-dependent cytotoxicity using mitochondrial MTT reduction assays, and the effective threshold concentration was in nanomolar level. At the concentrations less than 10 nM, ouabain induced a decrease of mitochondrial activity until a 7-hr exposure was performed, while it induced a rapid drop of mitochondrial function as early as a 2-hr treatment of cells with high concentrations of ouabain suggesting the involvement of two distinct mechanisms to ouabain action. After functional examinations, the data showed that both low and high concentrations of ouabain induced an inhibition of Na+-K+ ATPase and a subsequent Ca-45(2+) influx into PC-3 cells. High concentrations of ouabain induced a significant and time-dependent loss of mitochondrial membrane potential (A,,,), a sustained production of reactive oxygen species (ROS), and severe apoptotic reaction. Ouabain also induced an increase of Par-4 (prostate apoptosis response 4) expression. Furthermore, an antisense, but not nonsense, oligomer against Par-4 expression significantly inhibited the cytotoxicity induced by low concentrations of ouabain. It is suggested that ouabain induces two modes of cytotoxic effect in human hormone-independent prostate cancer PC-3 cells. Low concentrations of ouabain induce the increase of Par-4 expression and sensitize the cytotoxicity; while high concentrations of ouabain induce a loss of DeltaPsi(m), a sustained ROS production and a severe apoptosis in PC-3 cells. (C) 2003 Elsevier Inc. All rights reserved.