AbstractChoroidal neovascularization (CNV), is a major cause of irreversible blindness among the elderly population in developed countries, which is resulted from subretinal fibrosis without effective therapeutic strategies. Endothelial‐to‐mesenchymal transition (EndMT) of choroidal vascular endothelial cells (CVECs) contributes to subretinal fibrosis. Lycopene (LYC), a non‐pro‐vitamin A carotenoid, plays an anti‐fibrotic role. Herein, we explored the effect and mechanism of LYC on the EndMT of CVECs during CNV. Firstly, LYC inhibited EndMT in hypoxic human choroidal endothelial cells (HCVECs). Meanwhile, LYC inhibited proliferation, androgen receptor (AR) expression and nuclear localization in hypoxic HCVECs. Then LYC‐inhibited AR promotes the activation of microphthalmia‐associated transcription factor (MITF) in hypoxic HCVECs. In addition, LYC down‐regulated AR and induced MITF up‐regulated pigment epithelium‐derived factor (PEDF) transcription and expression in hypoxic HCVECs. Moreover, LYC‐induced PEDF bound to laminin receptor (LR), inhibiting EndMT of hypoxic HCVECs via down‐regulating protein kinase B (AKT)/β‐catenin pathway. In vivo, LYC alleviated mouse laser‐induced subretinal fibrosis secondary to CNV via up‐regulating PEDF without any ocular or systemic toxicity. These results indicate that LYC inhibits EndMT of CVECs via modulating AR/MITF/PEDF/LR/AKT/β‐catenin pathway, showing LYC is a promising therapeutic agent for CNV.