c-Src regulates cell cycle proteins expression through protein kinase B/glycogen synthase kinase 3 beta and extracellular signal-regulated kinases 1/2 pathways in MCF-7 cells
c-Src regulates cell cycle proteins expression through protein kinase B/glycogen synthase kinase 3 beta and extracellular signal-regulated kinases 1/2 pathways in MCF-7 cells
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c-Src 通过蛋白激酶 B/糖原合酶激酶 3 beta 和细胞外信号调节激酶 1/2 途径调节 MCF-7 细胞中的细胞周期蛋白表达
DOI：
10.1093/abbs/gmt042
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发表时间：
2013-07-01
影响因子：
3.7
通讯作者：
Feng, Renqing
中科院分区：
文献类型：
--
作者：
Liu, Xiang;Du, Liying;Feng, Renqing
我们已经证明，c-Src抑制抑制人乳腺癌细胞的上皮间质转化。在此，我们使用siRNA和小分子抑制剂4-氨基-5-（4-氯苯基）-7-（叔丁基）吡唑并[3，4-d]嘧啶（PP 2）研究了c-Src在细胞周期进程中的作用。蛋白质印迹分析表明，PP 2抑制c-Src后，细胞周期蛋白D1和细胞周期蛋白E下调，p27 Kip 1上调。在PP 2存在下孵育细胞显著阻断细胞外信号调节激酶1/2（ERK 1/2）、蛋白激酶B（AKT）和糖原合成酶激酶3 β（GSK 3）的磷酸化。使用MEK 1/2/ERK 1/2和磷脂酰肌醇3-激酶/AKT通路的特异性药理学抑制剂来证明信号级联与细胞周期蛋白表达之间的关系。抑制ERK 1/2或AKT活性后，细胞周期蛋白D1和细胞周期蛋白E的表达下降，而p27 Kip 1的表达增加。此外，通过siRNA敲低c-Src降低了细胞增殖和ERK 1/2、AKT和GSK 3的磷酸化。在c-Src被siRNA耗尽后，我们观察到细胞周期蛋白D1和细胞周期蛋白E的显著下调，以及p27 Kip 1的上调。这些结果表明，PP 2或siRNA抑制c-Src可能通过AKT/GSK 3和ERK 1/2途径调节细胞周期的进展。
We have demonstrated that c-Src suppression inhibited the epithelial to mesenchymal transition in human breast cancer cells. Here, we investigated the role of c-Src on the cell cycle progression using siRNAs and small molecule inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2). Western blot analysis demonstrated the down-regulation of cyclin D1 and cyclin E and up-regulation of p27 Kip1 after c-Src suppression by PP2. Incubation of cells in the presence of PP2 significantly blocked the phosphorylation of extracellular signal-regulated kinases 1/2 (ERK1/2), protein kinase B (AKT), and glycogen synthase kinase 3 beta (GSK3). Specific pharmacological inhibitors of MEK1/2/ERK1/2 and phosphatidylinositide 3-kinase/AKT pathways were used to demonstrate the relationship between the signal cascade and cell cycle proteins expression. The expression of cyclin D1 and cyclin E were decreased after inhibition of ERK1/2 or AKT activity, whereas the p27 Kip1 expression was increased. In addition, knockdown of c-Src by siRNAs reduced cell proliferation and phosphorylation of ERK1/2, AKT, and GSK3. After c-Src depletion by siRNAs, we observed significant down-regulation of cyclin D1 and cyclin E, and up-regulation of p27 Kip1. These results suggest that c-Src suppression by PP2 or siRNAs may regulate the progression of cell cycle through AKT/GSK3 and ERK1/2 pathways.