miR-140-5p suppresses retinoblastoma cell proliferation, migration, and invasion by targeting CEMIP and CADM3

Cell Mol Biol (Noisy-le-grand). 2018 May 15;64(6):42-47.

Abstract

Retinoblastoma (RB) is a childhood intraocular tumor, affecting millions of patients worldwide. MicroRNA-140-5p (miR-140-5p) was demonstrated to be involved in the tumorigenesis of various human cancers; however, its role in RB remains undetermined. In this study, quantitative real-time PCR (qRT-PCR) and Western blot assays were used to determine the expression levels of miR-140-5p, cell migration-inducing protein (CEMIP), and cell adhesion molecule 3 (CADM3) in RB tissues and cell-lines. The proliferation ability was detected by cell-counting kit 8 (CCK-8), Edu staining, and colony formation assay. The cell cycle and migration and invasion abilities were measured by flow cytometry, wound-healing assay and Transwell assays, respectively. The correlation between miR-140-5p and CEMIP/CADM3 were then confirmed by immunofluorescence (IF) and dual-luciferase reporter assays. The results showed that miR-140-5p expression was significantly decreased; however, CEMIP and CADM3 expression was increased in RB tissues and cells. Overexpression of miR-140-5p inhibited proliferation, migration, and invasion of RB cells. We also found that miR-140-5p inhibited CEMIP and CADM3 expressions in RB cells. In addition, we demonstrated that miR-140-5p might negatively regulate the transcriptional activities of CEMIP and CADM3 by targeting their 3'-UTR. Therefore, we suggested that miR-140-5p could be a potential therapeutic target for the treatment of RB through CEMIP and CADM3.

Keywords: Cell adhesion molecule 3; Cell migration inducing protein; MicroRNA-140-5p; Migration and invasion.; Proliferation; Retinoblastoma.

MeSH terms

  • Cell Adhesion Molecules / antagonists & inhibitors*
  • Cell Adhesion Molecules / metabolism
  • Cell Division
  • Cell Line, Tumor
  • Cell Movement
  • Eye Neoplasms / genetics
  • Eye Neoplasms / pathology*
  • Gene Expression Regulation, Neoplastic*
  • Genes, Reporter
  • Humans
  • Hyaluronoglucosaminidase
  • Immunoglobulins / metabolism
  • MicroRNAs / genetics
  • MicroRNAs / physiology*
  • Molecular Targeted Therapy
  • Neoplasm Invasiveness
  • Neoplasm Proteins / antagonists & inhibitors
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / physiology*
  • Proteins / antagonists & inhibitors*
  • Proteins / metabolism
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • RNA, Neoplasm / biosynthesis
  • RNA, Neoplasm / genetics
  • RNA, Neoplasm / physiology*
  • Recombinant Proteins / metabolism
  • Retinoblastoma / genetics
  • Retinoblastoma / pathology*
  • Tumor Cells, Cultured
  • Tumor Stem Cell Assay

Substances

  • CADM3 protein, human
  • Cell Adhesion Molecules
  • Immunoglobulins
  • MicroRNAs
  • Mirn140 microRNA, human
  • Neoplasm Proteins
  • Proteins
  • RNA, Messenger
  • RNA, Neoplasm
  • Recombinant Proteins
  • CEMIP protein, human
  • Hyaluronoglucosaminidase