Javanmoghadam-Kamrani Sonia, Keyomarsi Khandan
Department of Experimental Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
Cell Cycle. 2008 Aug;7(15):2434-40. doi: 10.4161/cc.6364. Epub 2008 May 29.
Synchronization by Lovastatin arrests many cell types reversibly in the G(1) phase of the cell cycle. Here we show that Lovastatin (10 microM) 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.
洛伐他汀介导的同步化作用可使许多细胞类型在细胞周期的G(1)期发生可逆性停滞。在此我们表明,洛伐他汀(10微摩尔)可介导人乳腺癌细胞MCF-7和MDA-MB-231的细胞周期停滞,其中85%的细胞积聚在细胞周期的G(1)期。添加甲羟戊酸(浓度为洛伐他汀的100倍)可使细胞从G(1)期停滞中释放出来,并允许其同步进入细胞周期的G(1)晚期、S期和G(2)/M期。通过蛋白质印迹分析检测不同细胞周期蛋白的表达,作为细胞周期不同阶段的标志物,这表明在初始G(1)期停滞之后,细胞可同步过渡到每个细胞周期阶段。由于其同步水平以及同步细胞群体的高产量,洛伐他汀细胞同步化方法可用于检测多种不同细胞系中的基因表达模式。