Selective induction of apoptosis in mutant p53 premalignant and malignant cancer cells by PRIMA-1 through the c-Jun-NH2-kinase pathway
Selective induction of apoptosis in mutant p53 premalignant and malignant cancer cells by PRIMA-1 through the c-Jun-NH2-kinase pathway
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DOI：
10.1158/1535-7163.mct-04-0206
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发表时间：
2005-06-01
影响因子：
5.7
通讯作者：
Fine, RL
中科院分区：
文献类型：
--
作者：
Li, Y;Mao, Y;Fine, RL
PRIMA, 1 （p53重激活和诱导大量凋亡）是一种化合物，最初通过筛选低分子量化合物库被确定为选择性突变型p53依赖性生长抑制剂。然而，其作用机制尚不清楚。在本研究中，我们检测了PRIMA-1对三种人结直肠癌前腺瘤细胞系（RG/C2、BR/C1和AA/C1）和四种结直肠癌细胞系（DLD-1、SW480、LOVO和HCT116）的毒性及其作用机制。它只选择性地诱导突变型p53癌前和恶性结肠癌细胞系的凋亡，而对野生型p53癌前和恶性结肠癌细胞系没有毒性。利用温度敏感型p53突变体Ala 143的稳定转染剂在无p53 h1299肺癌细胞中，我们发现PRIMA-1在突变型p53构象（37℃）细胞中诱导的凋亡明显多于野生型p53构象（32.5℃）细胞。细胞周期分析表明其对细胞生长的抑制作用与诱导G(2)阻滞有关。Western blot分析显示，PRIMA-1选择性地增加了p53突变细胞中p21和GADD45的表达。然而，Fas、Bcl-2家族蛋白和caspases与PRIMA-1诱导的细胞死亡无关。c- jun - nh2激酶（JNK）抑制剂SP 600125，而不是p38丝裂原活化蛋白激酶抑制剂SB 203580或细胞外信号调节激酶抑制剂PD 98059，可以阻断PRIMA-1诱导的细胞凋亡。转染显性阴性磷酸化突变体JNK，而非显性阴性p38或野生型JNK，可以抑制PRIMA-1诱导的细胞死亡，这表明JNK途径在PRIMA-1诱导的细胞凋亡中起重要作用。PRIMA-1是一种对p53突变细胞具有高选择性毒性的小分子，可以作为开发新的p53靶向药物治疗癌前和恶性细胞的原型。
PRIMA, 1 (p53 reactivation and induction of massive apoptosis) is a chemical compound that was originally identified as a selective mutant p53-dependent growth suppressor by screening a library of low-molecular-weight compounds. However, its mechanism of action is unknown. In this study, we examined toxicity of PRIMA-1 to three premalignant human colorectal adenoma cell lines (RG/C2, BR/C1, and AA/C1) and four colorectal carcinoma cell lines (DLD-1, SW480, LOVO, and HCT116) and its mechanism of action. It selectively induced apoptosis only in the mutant p53 premalignant and malignant colon cell lines, but was not toxic to the wild-type p53 premalignant and malignant colon cell lines. Using stable transfectants of temperature-sensitive p53 mutant Ala 143 in null p53 H 1299 lung cancer cells, we found that PRIMA-1 induced significantly more apoptosis in cells with mutant p53 conformation (37 degrees C) than the wild-type p53 conformation (32.5 degrees C). Cell cycle analysis indicated that its inhibition of cell growth was correlated with induction of G(2) arrest. Western blot analysis showed PRIMA-1 increased p21 and GADD45 expression selectively in the mutant p53 cells. However, Fas, Bcl-2 family proteins, and caspases were not involved in PRIMA-1 -induced cell death. The c-Jun-NH2-kinase (JNK) inhibitor SP 600125, but not p38 mitogen-activated protein kinase inhibitor SB 203580 or extracellular signal-regulated kinase inhibitor PD 98059, blocked PRIMA-1 -induced apoptosis. Transfection with a dominant-negative phosphorylation mutant JNK, but not a dominant-negative p38 or wild-type JNK, inhibited PRIMA-1 -induced cell death, suggesting that the JNK pathway plays an important role in PRIMA-1 -induced apoptosis. PRIMA-1 is a highly selective small molecule toxic to p53 mutant cells and may serve as a prototype for the development of new p53-targeting agents for therapy of premalignant and malignant cells.