Yerly-Motta V, Pavy J J, Hervé P
Cellular Immunology Laboratory, ETS Franche-Comté, Besançon, France.
Biotech Histochem. 1999 May;74(3):119-28. doi: 10.3109/10520299909047963.
To obtain different cell populations at specific cell cycle stages, we used a cell culture synchronization protocol. Effects of five different cell cycle inhibitors acting throughout the cell cycle were examined by DNA flow cytometric analysis of a synchrony/release lymphoma cell line (CEM). The screening synchronized protocol showed that staurosporine, mimosine and aphidicolin are reversible G1 phase inhibitors that act at different times. Staurosporine acted in early G1, exhibited the strongest cytotoxic effect, and induced apoptosis. Mimosine and aphidicolin acted in late G1 and at the G1/S boundary, respectively. Hydroxyurea arrested CEM cells in early S phase, but later than the aphidicolin arrest point. Nocodazole synchronized CEM cells in M phase. All the inhibitors examined in this study can be used to synchronize cells at different phases of the cell cycle and were reversible with little toxicity except for staurosporine which is highly toxic. Because the regulatory mechanism of the cell cycle is disrupted by their effects on protein synthesis, however, these drugs must be used with caution.
为了在特定细胞周期阶段获得不同的细胞群体，我们使用了细胞培养同步化方案。通过对同步化/释放淋巴瘤细胞系（CEM）进行DNA流式细胞术分析，研究了作用于整个细胞周期的五种不同细胞周期抑制剂的效果。筛选同步化方案表明，星形孢菌素、含羞草碱和阿非迪霉素是在不同时间起作用的可逆G1期抑制剂。星形孢菌素作用于G1早期，表现出最强的细胞毒性作用，并诱导细胞凋亡。含羞草碱和阿非迪霉素分别作用于G1晚期和G1/S边界。羟基脲使CEM细胞停滞在S期早期，但比阿非迪霉素的停滞点晚。诺考达唑使CEM细胞同步于M期。本研究中检测的所有抑制剂均可用于使细胞在细胞周期的不同阶段同步化，除了毒性很强的星形孢菌素外，其他抑制剂均可逆转且毒性很小。然而，由于这些药物对蛋白质合成的影响会破坏细胞周期的调节机制，因此必须谨慎使用。
