G1 cell cycle arrest and apoptosis are induced in NIH 3T3 cells by KN-93, an inhibitor of CaMK-II (the multifunctional Ca2+/CaM kinase).
G1 cell cycle arrest and apoptosis are induced in NIH 3T3 cells by KN-93, an inhibitor of CaMK-II (the multifunctional Ca2+/CaM kinase).
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发表时间：
1995-09
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通讯作者：
Robert M. Tombes;Steven Grant;EH Westin;Geoffrey W. Krystal
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作者：
Robert M. Tombes;Steven Grant;EH Westin;Geoffrey W. Krystal
钙调蛋白-II(CaMK-II)是一种广泛存在的丝氨酸/苏氨酸蛋白激酶，由钙和钙调蛋白(CaM)激活，参与细胞周期调控。NIH 3T3成纤维细胞胞浆提取物包含CaMK-II酶活性和两种主要的钙/CaM依赖的磷酸蛋白M(R)55000和115,000。逆转录-聚合酶链式反应表明，CaMK-II的Gamma B和Gamma C同工酶主要表达。KN-93是一种新型的经膜途径的纯化的神经元CaMK-II的合成抑制剂，它对血清诱导的成纤维细胞生长的抑制作用与其对CaMK-II活性的抑制作用具有剂量依赖关系。KN-93作用2天后，95%的细胞滞留在G1期。G1期停滞是可逆的；KN-93释放后第1天，细胞高峰进入S和G2-M期。KN-92是一种类似的非活性化合物，对CaMK-II的活性和细胞生长没有影响。KN-93还阻断碱性成纤维细胞生长因子、血小板衍生生长因子-BB、表皮生长因子和胰岛素样生长因子-1刺激的细胞生长。KN-93诱导G1期停滞3d后，细胞体积缩小，存活率下降，DNA片段化，表明细胞发生了凋亡。这些数据表明，CaMK-II对细胞周期通过G1是必需的，并且在生长和/或生存因子信号转导的共同位置起作用。
CaMK-II (the type II multifunctional Ca2+/calmodulin kinase) is a ubiquitous serine/threonine protein kinase that is activated by Ca2+ and calmodulin (CaM) and has been implicated in cell cycle control. NIH 3T3 fibroblast cytosolic extracts contain CaMK-II enzymatic activity and two major Ca2+/CaM-dependent phosphoproteins of M(r) 55,000 and 115,000. Reverse transcription-PCR indicates that the gamma B and gamma C isozymes of CaMK-II are predominately expressed. KN-93, a novel membrane-permeant synthetic inhibitor of purified neuronal CaMK-II, inhibits serum-induced fibroblast cell growth in a comparable dose-dependent fashion to its inhibition of CaMK-II activity. After 2 days of KN-93 treatment, 95% of cells are arrested in G1. G1 arrest is reversible; 1 day after KN-93 release, a peak of cells had progressed into S and G2-M. KN-92, a similar but inactive compound, had no effect on CaMK-II activity or cell growth. KN-93 also blocked cell growth stimulated by basic fibroblast growth factor, platelet-derived growth factor-BB, epidermal growth factor, and insulin-like growth factor-1. After 3 days of KN-93-induced G1 arrest, cell size and viability decreased and DNA fragmented, indicating apoptosis. These data suggest that CaMK-II is necessary for cell cycle progression through G1 and operates at a site common to the transduction of signals from growth and/or survival factors.