The Role and Potential Mechanism of Nuclear Protein PCNP in Retinal Neovascularization in Vitro

Abstract Purpose Retinal neovascularization (RNV) drives vision loss in diseases like diabetic retinopathy, and current anti-VEGF therapies have limitations. PEST-containing nuclear protein (PCNP) has roles in malignancy and angiogenesis, but its function in RNV is unclear. This study explored the function and underlying mechanisms of PCNP in RNV under in vitro conditions. Methods To achieve PCNP overexpression or knockdown, human umbilical vein endothelial cells (HUVECs) were subjected to transfection. The effects on cell proliferation (CCK-8 assay), migration (wound healing assay), and in vitro angiogenesis (tube formation assay) were subsequently evaluated. VEGF levels in supernatants (ELISA, CBA) and cell lysates (Western blot) were quantified. Transcriptome sequencing was performed on PCNP-overexpressing HUVECs to identify altered signaling pathways. Results PCNP overexpression significantly inhibited HUVEC proliferation, migration, and tube formation, key steps in neovascularization. Conversely, PCNP knockdown promoted these processes. While supernatant VEGF decreased unexpectedly in both groups, intracellular VEGF levels remained unchanged. Transcriptome analysis highlighted enrichment of IL-17, TNF, and NF-κB inflammatory signaling pathways with PCNP overexpression. Conclusion PCNP inhibits endothelial cell processes essential for neovascularization in vitro. The mechanism appears potentially independent of direct VEGF modulation and may involve inflammatory signaling pathways. PCNP emerges as a novel factor in RNV regulation, warranting further investigation for potential therapeutic targeting.