Simultaneous suppression of cdc2 and cdk2 activities induces neuronal differentiation of PC12 cells
Simultaneous suppression of cdc2 and cdk2 activities induces neuronal differentiation of PC12 cells
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DOI：
10.1074/jbc.275.17.12572
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发表时间：
2000-04-28
影响因子：
4.8
通讯作者：
Kameya, T
中科院分区：
文献类型：
--
作者：
Dobashi, Y;Shoji, M;Kameya, T
检测了cdc 2和cdk 2在大鼠嗜铬细胞瘤PC 12细胞神经元分化过程中的参与。当PC 12细胞与神经生长因子（NGF）共同培养时，cdc 2的表达在第5天后显著下降，而cdk 2的表达在第7天后逐渐下降。过表达cdc 2或cdk 2的细胞对NGF诱导的分化和生长抑制具有抗性，并在NGF处理期间分别保持高的cdc 2或cdk 2激酶活性。与此相反，神经生长因子处理的亲本细胞显示出显着下降，这些激酶活性后3天。当用cdc 2/cdk 2的特异性抑制剂（丁内酯-I，olomoucin）处理PC 12细胞时，它们表现出显著的轴突延伸和微管相关蛋白2表达的上调。另外，用cdc 2和cdk 2的反义寡核苷酸混合物处理导致cdc 2和cdk 2激酶活性的下调，以及显著的轴突生长和微管相关蛋白2表达的上调。然而，在用单一反义寡核苷酸或反义cdc 2 + cdk 4或cdk 2 + cdk 4寡核苷酸混合物处理的细胞中未观察到神经突生长。这些结果表明，同时下调cdc 2和cdk 2的活性是足够的，在PC 12细胞的神经元分化所必需的。
The involvement of cdc2 and cdk2 during neuronal differentiation in rat pheochromocytoma PC12 cells was examined. When PC12 cells were cultured with nerve growth factor (NGF), expression of cdc2 decreased significantly after day 5, while expression of cdk2 decreased gradually after day 7. Cells overexpressing cdc2 or cdk2 were resistant to NGF-induced differentiation and growth suppression, and maintained high cdc2 or cdk2 kinase activity, respectively, during NGF treatment. In contrast, the NGF-treated parental cells showed a marked decline in these kinase activities after day 3. When PC12 cells were treated with specific inhibitors of cdc2/cdk2 (butyrolactone-I, olomoucin), they showed marked neurite extension and up-regulation of microtubule-associated protein 2 expression. In addition, treatment with mixtures of antisense oligonucleotides for cdc2 and cdk2 resulted in down-regulation of both cdc2 and cdk2 kinase activities as well as significant neurite outgrowth and up-regulation of microtubule-associated protein 2 expression. However, neurite outgrowth was not observed in cells treated with either single antisense oligonucleotide, or antisense cdc2 + cdk4 or cdk2 + cdk4 oligonucleotide mixtures. These results suggest that simultaneous down-regulation of cdc2 and cdk2 activity is sufficient and necessary for neuronal differentiation in PC12 cells.