miR-4529-3p Promotes the Progression of Retinoblastoma by Inhibiting RB1 Expression and Activating the ERK Signaling Pathway

Mol Biotechnol. 2024 Jan;66(1):102-111. doi: 10.1007/s12033-023-00738-7. Epub 2023 Apr 11.

Abstract

Retinoblastoma (RB) is a malignant ocular cancer that affects children. Several microRNAs (miRNAs) have been implicated in RB regulation. The present study aimed to investigate the role of miR-4529-3p in RB pathogenesis. Scratch, Transwell, and Cell Counting Kit (CCK)-8 assays were conducted to assess the migratory, invasive, and proliferative abilities of RB cells. The expression levels of miR-4529-3p, RB1, and ERK pathway-related proteins were analyzed using western blotting and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Target relationships were verified using dual-luciferase reporter experiments. A murine RB model was developed to analyze the effects of miR-4529-3p on RB tumor growth in vivo. Our experiments revealed high levels of miR-4529-3p and low levels of RB1 in RB tissues. Functional analyses revealed that the migratory, invasive, and proliferative abilities of RB cells were repressed by miR-4529-3p inhibition. Similarly, p-ERK 1/2 protein levels were suppressed by miR-4529-3p inhibition. Furthermore, downregulation of miR-4529-3p limited tumor growth in vivo. Mechanistically, miR-4259-3p targets RB1. Interestingly, RB1 silencing abrogated the alleviative effects of miR-4529-3p downregulation in RB cells. MiR-4529-3p promotes RB progression by inhibiting RB1 and activating the ERK pathway. This evidence suggests that the miR-4529-3p/RB1 regulatory axis may be a prospective target for RB treatment in clinical settings.

Keywords: ERK pathway; RB1; Retinoblastoma; miR-4529-3p.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Cell Proliferation / genetics
  • Child
  • Gene Expression Regulation, Neoplastic
  • Humans
  • MAP Kinase Signaling System / genetics
  • Mice
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Retinal Neoplasms* / genetics
  • Retinal Neoplasms* / metabolism
  • Retinal Neoplasms* / pathology
  • Retinoblastoma Binding Proteins / genetics
  • Retinoblastoma Binding Proteins / metabolism
  • Retinoblastoma* / genetics
  • Retinoblastoma* / metabolism
  • Retinoblastoma* / pathology
  • Signal Transduction
  • Ubiquitin-Protein Ligases / metabolism

Substances

  • MicroRNAs
  • RB1 protein, human
  • Ubiquitin-Protein Ligases
  • Retinoblastoma Binding Proteins