NCC-RbC-T1

Background

Retinoblastoma (Rb) is the most common intraocular cancer in children. This study examined netrin-1 expression and function using public databases, clinical specimens, and cell line models. Netrin-1 was significantly upregulated in Rb. Silencing netrin-1 with siRNA did not affect Rb cell proliferation but reduced HUVEC vessel formation in vitro. In nude mice, netrin-1 knockdown suppressed tumor growth and vessel density, and VEGF-A co-expression reversed these effects. These findings suggest netrin-1 regulates Rb-associated angiogenesis and may be a therapeutic target.

Key Findings

Netrin-1 was upregulated in Rb samples and cell lines Y79, WERI-Rb-1, NCC-Rbc-57, NCC-Rbc-92, and NCC-Rbc-T1 compared to normal retina, confirmed by immunohistochemistry, RT-qPCR and ELISA. Bioinformatics of GEO datasets showed angiogenesis pathway enrichment and netrin-1 upregulation. siRNA-mediated netrin-1 silencing effectively reduced mRNA and protein levels. In vitro, netrin-1 depletion did not alter Rb cell growth by CCK-8 but significantly decreased HUVEC vessel structure formation in co-culture. In vivo, subcutaneous WERI-Rb-1 xenografts with netrin-1 knockdown showed reduced tumor growth by bioluminescence and lower CD31+ vessel density. Administration of recombinant VEGF-A reversed the suppression of tumor growth and vessel density, linking netrin-1 effects to VEGF-A-dependent angiogenesis. These data indicate netrin-1 promotes Rb-associated angiogenesis and tumor progression, representing a potential therapeutic target.

Contribution

Creative Bioarray provided human retinoblastoma cell lines NCC-Rbc-57, NCC-Rbc-92, and NCC-Rbc-T1 used in this study. These cell lines enabled netrin-1 expression analysis, siRNA silencing validation, and in vitro angiogenesis assays, supporting the conclusion that netrin-1 promotes Rb-associated angiogenesis and tumor progression.