Pifithrin-α supresses p53 and protects cochlear and vestibular hair cells from cisplatin-induced apoptosis
Pifithrin-α supresses p53 and protects cochlear and vestibular hair cells from cisplatin-induced apoptosis
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DOI：
10.1016/s0306-4522(03)00286-0
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发表时间：
2003-08
期刊：
影响因子：
3.3
通讯作者：
M. Zhang;W. Liu;Dalian Ding;Richard Salvi
中科院分区：
文献类型：
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作者：
M. Zhang;W. Liu;Dalian Ding;Richard Salvi
顺铂是一种常用的麻醉剂，它会破坏耳蜗和前庭系统中的感觉毛细胞，导致不可逆的听力损失和平衡问题。顺铂诱导的毛细胞损伤可能通过细胞凋亡发生。最近的研究表明，p53可能在启动顺铂诱导的某些细胞类型的凋亡中发挥重要作用。为了确定p53是否在顺铂介导的毛细胞损失中起作用，从出生后第3-4天的大鼠制备耳蜗和椭圆囊器官型培养物，并用顺铂或顺铂加pifithrin-α（PFT）（一种p53抑制剂）处理。对照培养物缺乏p53免疫标记、半胱天冬酶-1和半胱天冬酶-3标记，并且蛋白质印迹中不存在p53蛋白。顺铂（1-10 μg/ml）引起耳蜗和椭圆囊培养物中毛细胞的剂量依赖性损失，上调磷酸化p53丝氨酸15免疫标记，增加顺铂治疗开始后6 - 48 h Western印迹中磷酸化p53丝氨酸15的表达，并增加耳蜗和前庭培养物中caspase-1和caspase-3的标记。在顺铂处理的耳蜗和椭圆囊培养物中加入PFT（20-100 μM）导致毛细胞存活率呈剂量依赖性增加;抑制蛋白质印迹中p53的表达，并消除培养物中的caspase-1和caspase-3标记。这些结果表明，肿瘤抑制蛋白p53在启动耳蜗和前庭毛细胞凋亡中发挥着关键作用。用PFT暂时抑制p53提供了对顺铂诱导的毛细胞损失的显著保护，并提供了减少顺铂的耳毒性、前庭毒性和神经毒性副作用的可能性。
Cisplatin, a commonly used antineoplastic agent, destroys the sensory hair cells in the cochlear and vestibular system leading to irreversible hearing loss and balance problems. Cisplatin-induced hair cell damage presumably occurs by apoptosis. Recent studies suggest that p53 may play an important role initiating cisplatin-induced apoptosis in some cell types. To determine if p53 plays a role in cisplatin-mediated hair cell loss, cochlear and utricular organotypic cultures were prepared from postnatal day 3–4 rats and treated with cisplatin or cisplatin plus pifithrin-α (PFT), a p53 inhibitor. Control cultures were devoid of p53 immunolabeling, caspase-1 and caspase-3 labeling and p53 protein was absent from Western blots. Cisplatin (1–10 μg/ml) caused a dose-dependent loss of hair cells in cochlear and utricular cultures, up-regulated phospho-p53 serine 15 immunolabeling, increased the expression of phospho-p53 serine 15 in Western blots from 6 to 48 h after the onset of cisplatin-treatment, and increased caspase-1 and caspase-3 labeling in cochlear and vestibular cultures. Addition of PFT (20–100 μM) to cisplatin-treated cochlear and utricular cultures resulted in a dose-dependent increase in hair cell survival; suppressed the expression of p53 in Western blots and eliminated caspase-1 and caspase-3 labeling in cultures. These results suggest that the tumor suppressor protein, p53, plays a critical role in initiating apoptosis in cochlear and vestibular hair cells. Temporary suppression of p53 with PFT provides significant protection against cisplatin-induced hair cell loss and offers the potential for reducing the ototoxic, vestibulotoxic and neurotoxic side effects of cisplatin.