Ovarian cancer G protein coupled receptor 1 suppresses cell migration of MCF7 breast cancer cells via a Gα12/13-Rho-Rac1 pathway.
Ovarian cancer G protein coupled receptor 1 suppresses cell migration of MCF7 breast cancer cells via a Gα12/13-Rho-Rac1 pathway.
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DOI：
10.1186/1750-2187-8-6
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发表时间：
2013-05-10
影响因子：
--
通讯作者：
Sang J
中科院分区：
文献类型：
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作者：
Li J;Guo B;Wang J;Cheng X;Xu Y;Sang J
卵巢癌G蛋白偶联受体1（OGR 1）介导对人前列腺和卵巢癌细胞迁移的抑制作用。然而，介导这些抑制作用的机制和信号通路基本上是未知的。选择MCF 7细胞系作为模型系统来研究OGR 1调节细胞迁移的机制，因为它表达非常低水平的内源性OGR 1。使用伤口愈合和transwell迁移测定来评估细胞迁移活性。使用药理学抑制剂和相关基因的遗传形式以及小G蛋白下拉活性测定来研究所涉及的信号通路。通过Western blot和定量PCR分析各种信号分子的表达水平。OGR 1在MCF 7细胞中的过表达显著增强了Rho的激活和Rac 1的抑制，导致细胞迁移的抑制。此外，表达p115 RhoGEF的G蛋白信号传导（RGS）结构域的Gα12/13特异性调节因子，而不是用百日咳毒素（PTX，一种Gαi特异性抑制剂）处理，可以消除MCF 7细胞中OGR 1依赖的Rho激活、Rac 1失活和迁移抑制。测试的生物活性脂质对OGR 1在细胞迁移中的功能没有影响。我们的数据首次表明，OGR 1通过Gα12/13 -Rho-Rac 1信号通路抑制MCF 7细胞的细胞迁移。该途径不受生物活性脂质的显著影响，并且所有测定均在恒定pH下进行，表明OGR 1的组成型活性。这是第一次清楚地描绘了OGR 1介导的细胞信号通路参与迁移。
Ovarian cancer G protein coupled receptor 1 (OGR1) mediates inhibitory effects on cell migration in human prostate and ovarian cancer cells. However, the mechanisms and signaling pathways that mediate these inhibitory effects are essentially unknown. MCF7 cell line was chosen as a model system to study the mechanisms by which OGR1 regulates cell migration, since it expresses very low levels of endogenous OGR1. Cell migratory activities were assessed using both wound healing and transwell migration assays. The signaling pathways involved were studied using pharmacological inhibitors and genetic forms of the relevant genes, as well as small G protein pull-down activity assays. The expression levels of various signaling molecules were analyzed by Western blot and quantitative PCR analysis. Over-expression of OGR1 in MCF7 cells substantially enhanced activation of Rho and inhibition of Rac1, resulting in inhibition of cell migration. In addition, expression of the Gα12/13 specific regulator of G protein signaling (RGS) domain of p115RhoGEF, but not treatment with pertussis toxin (PTX, a Gαi specific inhibitor), could abrogate OGR1-dependent Rho activation, Rac1 inactivation, and inhibition of migration in MCF7 cells. The bioactive lipids tested had no effect on OGR1 function in cell migration. Our data suggest, for the first time, that OGR1 inhibits cell migration through a Gα12/13 -Rho-Rac1 signaling pathway in MCF7 cells. This pathway was not significantly affected by bioactive lipids and all the assays were conducted at constant pH, suggesting a constitutive activity of OGR1. This is the first clear delineation of an OGR1-mediated cell signaling pathway involved in migration.