Synchronization of the cell cycle using lovastatin
Synchronization of the cell cycle using lovastatin
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DOI：
10.4161/cc.6364
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发表时间：
2008-08-01
期刊：
影响因子：
4.3
通讯作者：
Keyomarsi, Khandan
中科院分区：
文献类型：
--
作者：
JavanMoghadam-Kamrani, Sonia;Keyomarsi, Khandan
通过洛伐他汀的同步化可逆地将许多细胞类型阻滞在细胞周期的G（1）期。在这里，我们表明，洛伐他汀（10 μ M）介导的人乳腺癌细胞，MCF-7和MDA-MB-231，其中85%的细胞积累在G（1）期的细胞周期停滞。加入甲羟戊酸（浓度为洛伐他汀的100倍）可使细胞从G（1）停滞状态释放，并允许同步进入细胞周期的晚期G（1）、S和G（2）/M期。通过蛋白质印迹分析检测作为细胞周期不同阶段的标志物的不同细胞周期蛋白的表达，并指示在初始G（1）停滞后同步过渡到每个细胞周期阶段。由于其同步水平和同步细胞群体的高产率，细胞同步化的洛伐他汀方法可用于检查各种不同细胞系中的基因表达模式。
Synchronization by Lovastatin arrests many cell types reversibly in the G(1) phase of the cell cycle. Here we show that Lovastatin (10 mu M) mediates cell cycle arrest in human breast cancer cells, MCF-7 and MDA-MB-231, where 85% of cells accumulate in the G(1) phase of the cell cycle. Addition of mevalonate (at 100X the Lovastatin concentration) releases the cells from the G(1) arrest and allows for synchronous entry into late G(1), S and G(2)/M phases of the cell cycle. The expressions of different cyclins as a marker for different phases of the cell cycle are detected by western blot analysis and indicative of synchronous transition into each of cell cycle phases following the initial G(1) arrest. Due to its level of synchrony and high yield of synchronous populations of cells, Lovastatin method of cell synchronization can be used for examining gene expression patterns in a variety of different cell lines.