Glycogen Synthase Kinase-3beta regulates Snail and beta-catenin during gastrin-induced migration of gastric cancer cells.
Glycogen Synthase Kinase-3beta regulates Snail and beta-catenin during gastrin-induced migration of gastric cancer cells.
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DOI：
10.1186/1750-2187-5-9
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发表时间：
2010-07-16
影响因子：
--
通讯作者：
Rana B
中科院分区：
文献类型：
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作者：
Mishra P;Senthivinayagam S;Rana A;Rana B
已知胃肠道肽激素胃泌素可调节胃肠道 (GI) 细胞的增殖、迁移和转移等多种细胞过程。本文描述的研究旨在详细阐明介导酰胺化胃泌素 (G17) 迁移反应的信号通路，并了解丝氨酸/苏氨酸激酶糖原合成酶激酶 3 beta (GSK3β) 在此过程中的参与。我们的结果表明，过表达 CCK2 受体的胃癌细胞（AGSE 细胞）与 G17 一起孵育会导致 GSK3βSer9 磷酸化的剂量和时间依赖性增加，表明激酶受到抑制。用 PI3 激酶途径的药理学抑制剂（渥曼青霉素）进行预处理无法拮抗 G17 诱导的 GSK3βSer9 磷酸化，表明这可能涉及 PI3 激酶独立途径。 G17 治疗还与 Snail 表达增加和 β-连环蛋白核转位有关，这两者都是 GSK3β 下游靶标。在 G17 不存在的情况下，用 GSK3β (AR-A014418) 药理学抑制剂进行预处理可增强 Snail 表达和 β-连环蛋白核转位，而 GSK3β (S9A) 磷酸化缺陷突变体的过度表达会消除 Snail 启动子诱导。这些表明 G17 通过抑制 GSK3β 来调节 Snail 和 β-catenin 通路。此外，GSK3β野生型（WT）或S9A突变体的过表达抑制了G17诱导的迁移和MMP7启动子诱导。在小干扰 RNA (siRNA) 介导的 Snail 和 β-catenin 表达敲低后设计的 G17 研究表明，在联合敲低两种蛋白后，G-17 诱导的迁移和 MMP7 启动子诱导显着减少。我们的研究表明，抑制 GSK3β 对于激活 G17 诱导的胃癌细胞迁移途径是必要的。 GSK3β 的抑制会导致 Snail 表达和 β-catenin 核转位的诱导，这两者都参与促进 G17 诱导的迁移。
The gastrointestinal peptide hormone gastrin is known to regulate various cellular processes including proliferation, migration and metastasis in gastrointestinal (GI) cells. The studies described here were undertaken to elucidate in detail the signaling pathways mediating the migratory responses of amidated gastrin (G17) and to understand the involvement of the serine/threonine kinase Glycogen Synthase Kinase-3 beta (GSK3β) in this. Our results indicate that incubation of gastric cancer cells overexpressing CCK2 receptor (AGSE cells) with G17 results in a dose and time dependent increase of GSK3βSer9 phosphorylation, indicative of an inhibition of the kinase. Pretreatment with a pharmacological inhibitor of PI3Kinase pathway (Wortmannin) was unable to antagonize G17-induced GSK3βSer9 phosphorylation, suggesting that this might involve PI3Kinase-independent pathways. Treatment with G17 was also associated with increased Snail expression, and β-catenin nuclear translocation, both of which are GSK3β downstream targets. Pretreatment with a pharmacological inhibitor of GSK3β (AR-A014418) augmented Snail expression and β-catenin nuclear translocation in the absence of G17, whereas overexpression of a phosphorylation deficient mutant of GSK3β (S9A) abrogated Snail promoter induction. These suggested that G17 modulates Snail and β-catenin pathways via inhibiting GSK3β. In addition, overexpression of GSK3β wild type (WT) or S9A mutant inhibited G17-induced migration and MMP7 promoter induction. G17 studies designed following small interference RNA (siRNA)-mediated knockdown of Snail and β-catenin expression indicated a significant reduction of G-17-induced migration and MMP7 promoter induction following combined knockdown of both proteins. Our studies indicate that inhibition of GSK3β is necessary to activate G17-induced migratory pathways in gastric cancer cells. Inhibition of GSK3β leads to an induction of Snail expression and β-catenin nuclear translocation, both of which participate to promote G17-induced migration.