Eo Jaejun, Kim Nahyun, Ghil Sungho
Department of Life Science, Kyonggi University, Suwon, Republic of Korea.
Department of Life Science, Kyonggi University, Suwon, Republic of Korea.
J Biol Chem. 2025 Aug 5;301(9):110554. doi: 10.1016/j.jbc.2025.110554.
Sphingosine-1-phosphate receptor 2 and 3 (S1P and S1P) are G protein-coupled receptors that mediate extracellular sphingosine-1-phosphate (S1P) signaling into cells. S1P and S1P are highly expressed in colon cancer cells, but their roles in cancer progression-related cellular phenotypes are not well understood. Recent studies suggest that regulator of G protein signaling 2 (RGS2) interacts with G protein-coupled receptors, either directly or indirectly, to regulate their signaling. However, the precise role of RGS2 in S1P and S1P signaling remains uninvestigated. In this study, we examined the interaction of RGS2 with S1P and S1P using bioluminescence resonance energy transfer analysis and assessed its impact on S1P- and S1P-mediated signaling in 293T and HCT116 cells. Bioluminescence resonance energy transfer analysis revealed that RGS2 and Gα subunits simultaneously bind to S1P and S1P. Furthermore, in the presence of these receptors, RGS2 translocated from the cytoplasm to the cell membrane. These interactions and membrane translocation were not observed with the RGS1 negative control, highlighting the specificity of RGS2 for S1P and S1P. RGS2 expression inhibited the activation of Gαi, Gαq, and Gα12, key signaling pathways mediated by S1P and S1P. S1P and S1P activation significantly enhanced cell migration and the expression of cancer-associated genes, effects that were effectively suppressed by RGS2 expression. In contrast, RGS1 failed to inhibit S1P- and S1P-mediated Gα signaling as well as downstream effects, such as enhanced cell migration and cancer-associated gene expression. Our findings show that RGS2 suppresses S1P2- and S1P3-mediated cancer-associated cellular phenotypes by interacting with these receptors and inhibiting Gα-mediated signaling.
1-磷酸鞘氨醇受体2和3（S1P₂和S1P₃）是G蛋白偶联受体，可将细胞外1-磷酸鞘氨醇（S1P）信号传导至细胞内。S1P₂和S1P₃在结肠癌细胞中高表达，但其在癌症进展相关细胞表型中的作用尚不清楚。最近的研究表明，G蛋白信号调节剂2（RGS2）直接或间接与G蛋白偶联受体相互作用，以调节其信号传导。然而，RGS2在S1P₂和S1P₃信号传导中的确切作用仍未得到研究。在本研究中，我们使用生物发光共振能量转移分析检测了RGS2与S1P₂和S1P₃的相互作用，并评估了其对293T和HCT116细胞中S1P₂和S1P₃介导的信号传导的影响。生物发光共振能量转移分析显示，RGS2和Gα亚基同时与S1P₂和S1P₃结合。此外，在这些受体存在的情况下，RGS2从细胞质转移到细胞膜。RGS1阴性对照未观察到这些相互作用和膜转位，突出了RGS2对S1P₂和S1P₃的特异性。RGS2表达抑制了Gαi、Gαq和Gα12的激活，这是S1P₂和S1P₃介导的关键信号通路。S1P₂和S1P₃激活显著增强细胞迁移和癌症相关基因的表达，而RGS2表达有效地抑制了这些效应。相比之下，RGS1未能抑制S1P₂和S1P₃介导的Gα信号传导以及下游效应，如增强的细胞迁移和癌症相关基因表达。我们的研究结果表明，RGS2通过与这些受体相互作用并抑制Gα介导的信号传导，抑制S1P₂和S1P₃介导的癌症相关细胞表型。
