AIM: To investigate the effects of collagen and opticin on the bioactivity of human retinal vascular endothelial cells (hRVECs), and explore its regulations by integrins and RhoA/ROCK1 signal pathway. METHODS: hRVECs were cultured in collagen and treated by opticin, and cell-based bioactivity assays of cell proliferation, migration, and adhesion were performed. The expression of integrin α2, integrin β1, RhoA and ROCK1 were examined with real-time PCR and Western blotting. RESULTS: Collagen could promote cell viability of proliferation and migration (all P<0.05), and enhance the mRNA expression of integrin α2, integrin β1, RhoA and ROCK1 (all P<0.05). Opticin could inhibit proliferation and migration ability of hRVECs cultured in collagen, and reduce the mRNA expression of integrin α2, integrin β1, RhoA and ROCK1 (all P<0.05). CONCLUSION: Collagen and opticin can affect bioactivity of hRVECs, which may be regulated by α2-, β1-integrins and RhoA/ROCK1 signal pathway.