CD97/ADGRE5, an adhesion G protein-coupled receptor (aGPCR), is highly expressed in several tumor cell types. CD97 has been shown to modulate tumorigenesis in part by promoting HUVEC migration, invasion and angiogenesis through the interaction with integrin alpha 5 beta 1 via its ectodomain RGD motif. In this study, we show that CD97 could induce angiogenesis via an alternative RGD-independent mechanism. Over-expression of CD97 with the wild-type or mutant RGD motif in HT1080 cells led to up-regulated MMP-9 and induced angiogenesis as revealed by the in vitro endothelial cell tube formation assay and in ovo chick chorioallantoic membrane assay. By contrast, expression of EMR2/ADGRE2, the CD97-homologous aGPCR that contains a corresponding SGD sequence, fails to induce angiogenesis due to lower MMP-9 expression. Interestingly, a single change of the SGD to RGD sequence allowed EMR2 to up-regulate MMP-9 expression, leading to enhanced angiogenesis. MMP-9 was shown to promote the proliferation, migration, and invasion of HUVEC partly by modulating the levels of VEGF, PIGF, and bFGF. Finally, we showed that the MMP-9 expression was in turn modulated by N-cadherin that was up-regulated by CD97 and EMR2/RGD. Our results indicate that two homologous aGPCRs, CD97 and EMR2, modulate angiogenesis and HUVEC proliferation, migration, and invasion through N-cadherin-regulated MMP-9 expression by RGD-independent and -dependent mechanisms, respectively. (C) 2019 Elsevier Inc. All rights reserved.