Regulator of G protein signaling 2 as a suppressor of sphingosine-1-phosphate 2- and 3-mediated signaling in colon cancer cells.

Sphingosine-1-phosphate receptor 2 and 3 (S1P(2) and S1P(3)) are G protein-coupled receptors that mediate extracellular sphingosine-1-phosphate (S1P) signaling into cells. S1P(2) and S1P(3) 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(2) and S1P(3) signaling remains uninvestigated. In this study, we examined the interaction of RGS2 with S1P(2) and S1P(3) using bioluminescence resonance energy transfer analysis and assessed its impact on S1P(2)- and S1P(3)-mediated signaling in 293T and HCT116 cells. Bioluminescence resonance energy transfer analysis revealed that RGS2 and Gα subunits simultaneously bind to S1P(2) and S1P(3). 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(2) and S1P(3). RGS2 expression inhibited the activation of Gαi, Gαq, and Gα12, key signaling pathways mediated by S1P(2) and S1P(3). S1P(2) and S1P(3) 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(2)- and S1P(3)-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.