Dual-agonist occupancy of orexin receptor 1 and cholecystokinin A receptor heterodimers decreases G-protein-dependent signaling and migration in the human colon cancer cell line HT-29
Dual-agonist occupancy of orexin receptor 1 and cholecystokinin A receptor heterodimers decreases G-protein-dependent signaling and migration in the human colon cancer cell line HT-29
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DOI：
10.1016/j.bbamcr.2017.03.003
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发表时间：
2017-07-01
影响因子：
5.1
通讯作者：
Chen, Jing
中科院分区：
文献类型：
--
作者：
Bai, Bo;Chen, Xiaoyu;Chen, Jing
食欲素（OX1R）和胆囊收缩素A （CCK1R）受体在人结肠癌细胞系HT-29的迁移过程中发挥相反的作用，可能参与了癌细胞侵袭转移的发病机制和病理生理过程。OX1R和CCK1R属于g蛋白偶联受体（gpcr）的A家族，但它们在实体瘤发展中的功能的详细机制尚不清楚。在这项研究中，我们研究了这两个受体是否异二聚，结果揭示了新的信号转导机制。生物发光和Forster共振能量转移以及接近连接实验表明，OX1R和CCK1R在HEK293和HT-29细胞中异二聚体的形成，并且OX1R跨膜结构域5对应的肽损害了异二聚体的形成。与单独使用orexin-A或CCK刺激相比，用orexin-A和CCK刺激OX1R和CCK1R异源二聚体可以降低G α G、G α i2、G α 12和G α 13的激活以及HT-29细胞的迁移，但没有改变GPCR与β -抑制素的相互作用。这些结果表明，OX1R和CCK1R异源二聚化在人结肠癌细胞中具有抗迁移作用。(C) 2017 Elsevier B.V.版权所有
The orexin (OX1R) and cholecystokinin A (CCK1R) receptors play opposing roles in the migration of the human colon cancer cell line HT-29, and may be involved in the pathogenesis and pathophysiology of cancer cell invasion and metastasis. OX1R and CCK1R belong to family A of the G-protein-coupled receptors (GPCRs), but the detailed mechanisms underlying their functions in solid tumor development remain unclear. In this study, we investigated whether these two receptors heterodimerize, and the results revealed novel signal transduction mechanisms. Bioluminescence and Forster resonance energy transfer, as well as proximity ligation assays, demonstrated that OX1R and CCK1R heterodimerize in HEK293 and HT-29 cells, and that peptides corresponding to transmembrane domain 5 of OX1R impaired heterodimer formation. Stimulation of OX1R and CCK1R heterodimers with both orexin-A and CCK decreased the activation of G alpha g, G alpha i2, G alpha 12, and G alpha 13 and the migration of HT-29 cells in comparison with stimulation with orexin-A or CCK alone, but did not alter GPCR interactions with beta-arrestins. These results suggest that OX1R and CCK1R heterodimerization plays an anti-migratory role in human colon cancer cells. (C) 2017 Elsevier B.V. All rights reserved.