Acetaminophen induces apoptosis of C6 glioma cells by activating the c-Jun NH(2)-terminal protein kinase-related cell death pathway.
Acetaminophen induces apoptosis of C6 glioma cells by activating the c-Jun NH(2)-terminal protein kinase-related cell death pathway.
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发表时间：
2001-10
影响因子：
3.6
通讯作者：
Myung-Ae Bae;J. Pie;Byoung Joon Song
中科院分区：
文献类型：
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作者：
Myung-Ae Bae;J. Pie;Byoung Joon Song
对乙酰氨基酚（AAP）是一种广泛使用的止痛药，大剂量服用会损害多种器官。在这项研究中，我们研究AAP是否通过改变与细胞死亡和存活相关的早期信号通路来引起细胞损伤。以C6胶质瘤细胞为模型，AAP诱导C6胶质瘤细胞凋亡和DNA片段化，呈时间和浓度依赖性。AAP在15 min内激活c-Jun N-末端蛋白激酶（JNK）5.3倍。JNK活性升高持续4 h，然后在8 h恢复到基础水平。相反，其他促分裂原活化蛋白（MAP）激酶的活性和细胞存活途径中Akt磷酸化水平在整个治疗过程中保持不变。磷脂酰肌醇-3激酶抑制剂渥曼青霉素或p38 MAP激酶抑制剂SB 203580不能降低AAP诱导的毒性，表明这些酶在细胞毒性中不起主要作用。AAP诱导的细胞凋亡之前的促凋亡Bax蛋白，细胞色素c的释放，和caspase-3活性的顺序升高。caspase抑制剂苄氧基羰基-Asp-Glu-Val-Asp-氟甲基酮（Z-DEVD-FMK）处理显著降低AAP诱导的caspase-3活化和细胞毒性。转染显性失活突变体JNK-KR或应激活化蛋白激酶激酶-1Lys->Arg突变体（SEK 1-KR）（JNK的直接上游激酶）的cDNA可显著降低AAP诱导的JNK活化和细胞死亡率。AAP的非细胞毒性类似物3-羟基乙酰苯胺既不增加JNK活性，也不引起细胞凋亡。用CYP 2 E1基因转录抑制剂YH 439预处理，显著降低CYP 2 E1 mRNA、蛋白含量和活性，以及AAP诱导的JNK激活和细胞死亡的速率。这些数据表明，AAP可以通过激活JNK相关的细胞死亡途径引起细胞损伤，为AAP诱导的细胞毒性提供了新的机制。
Acetaminophen (AAP), a widely used analgesic drug, can damage various organs when taken in large doses. In this study, we investigate whether AAP causes cell damage by altering the early signaling pathways associated with cell death and survival. AAP caused time- and concentration-dependent apoptosis and DNA fragmentation of C6 glioma cells used as a model. AAP activated c-Jun N-terminal protein kinase (JNK) by 5.3-fold within 15 min. The elevated JNK activity persisted for up to 4 h before it returned to the basal level at 8 h. In contrast, activities of other mitogen-activated protein (MAP) kinases and the level of Akt phosphorylation in the cell survival pathway remained unchanged throughout the treatment. Wortmannin, an inhibitor of phosphatidylinositol-3 kinase, or SB203580, an inhibitor of p38 MAP kinase, did not reduce AAP-induced toxicity, indicating that these enzymes do not play a major role in cell toxicity. AAP-induced apoptosis was preceded by the sequential elevation of the pro-apoptotic Bax protein, cytochrome c release, and caspase-3 activity. Treatment with caspase inhibitor benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethyl ketone (Z-DEVD-FMK) significantly reduced AAP-induced caspase-3 activation and cytotoxicity. Transfection of cDNA for the dominant-negative mutant JNK-KR or stress-activated protein kinase kinase-1 Lys-->Arg mutant (SEK1-KR), an immediate upstream kinase of JNK, significantly reduced AAP-induced JNK activation and cell death rate. The noncytotoxic analog of AAP, 3-hydroxyacetanilide, neither increased JNK activity nor caused apoptosis. Pretreatment with YH439, an inhibitor of CYP2E1 gene transcription, markedly reduced CYP2E1 mRNA, protein content, and activity, as well as the rate of AAP-induced JNK activation and cell death. These data indicate that AAP can cause cell damage by activating the JNK-related cell death pathway, providing a new mechanism for AAP-induced cytotoxicity.