Serum starvation and thymidine double blocking achieved efficient cell cycle synchronization and altered the expression of p27, p53, bcl-2 in canine breast cancer cells
Serum starvation and thymidine double blocking achieved efficient cell cycle synchronization and altered the expression of p27, p53, bcl-2 in canine breast cancer cells
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血清饥饿和胸苷双重阻断实现了有效的细胞周期同步并改变了犬乳腺癌细胞中p27、p53、bcl-2的表达
DOI：
10.1016/j.rvsc.2016.01.008
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发表时间：
2016-04-01
影响因子：
2.4
通讯作者：
Liu, Yun
中科院分区：
文献类型：
--
作者：
Tong, Jinjin;Sun, Dongdong;Liu, Yun
Cell synchronization is an approach to obtain cell populations of the same stage, which is a prerequisite to studying the regulation of cell cycle progression in vivo. Serum starvation and thymidine double blocking (TdR) are two important practices in studying cell cycle synchronization. However, their effects on canine cancer cells as well as the regulatory mechanisms by these two methods are poorly understood. In this study, we determined the optimum conditions of serum starvation and TdR and their effects on cell cycle synchronization. We further explored the involvement of PI3K/Akt signaling pathway in the cell cycle synchronization by investigating the expression of three key genes (p27, p53 and bcl-2). Serum starvation resulted in a reversible cell cycle arrest and synchronously progress through GO/G1. The highest percentage of CHMm cells (87.47%) in GO/G1 stage was obtained after 42 h incubation with 0.5% fetal bovine serum (FBS). TdR double blocking could arrest 98.9% of CHMm cells in G1/S phase (0 h of release), and could arrest 93.74% of CHMm cells in S phase after 4 h of release. We also found that the p27, p53, bcl-2 genes were most highly expressed in GO/G1 phase. Our current work revealed that serum starvation and TdR methods could achieve sufficient synchronization of CHMm cells. Moreover, the expression of p27, p53 and bcl-2 genes was related to cyclical movements and apoptosis. Our results will provide a new insight into cell cycle regulation and reprogramming of canine cancer cells induced by serum starvation and TdR blocking. (C) 2016 Elsevier Ltd. All rights reserved.
