Effects of miR-599 targeting YAP1 on proliferation, invasion and apoptosis of bladder urothelial carcinoma cells

Exp Mol Pathol. 2021 Feb:118:104599. doi: 10.1016/j.yexmp.2020.104599. Epub 2020 Dec 29.

Abstract

The aim of this study was to investigate the effects of microRNA (miR)-599 targeting Yes-associated protein 1 (YAP1) on the proliferation, invasion and apoptosis of bladder urothelial carcinoma cell line J82. J82 cells cultured in vitro were divided into miR-nc group (transfected with analog negative control miR-NC), miR-599 group (transfected with miR-599 analog), miR-599 + pcDNA3.1 group (co transfected with miR-599 analog and pcDNA3.1 empty vector), miR-599 + pcDNA3.1-YAP1 group (co transfected with miR-599 analog and pcDNA3.1-YAP1 over expression vector), the targeting relationship between miR-599 and YAP1 was detected by double luciferase reporter gene assay, the expression level of miR-599 was detected by real-time fluorescence quantitative PCR, the expression of YAP1 was detected by Western blot, cell proliferation was detected by MTT method, cell invasion was detected by Transwell's cell experiment, and apoptosis was detected by flow cytometry. The miR-599 was able to bind to YAP1 targetingly; compared with those in miR-NC group, miR-599 expression level and apoptosis rate in miR-599 group were significantly higher, however, the expression level of YAP1 protein and the abilities of cell proliferation and invasion were significantly lower (P < 0.05); there was no significant difference in the above indexes between miR-599 group and miR-599 + pcDNA3.1 group (P > 0.05); compared with those in miR-599 + pcDNA3.1 group, the expression level of YAP1 protein and the abilities of cell proliferation and invasion in miR-599 + pcDNA3.1-YAP1 group were significantly higher, while the apoptotic rate was significantly lower (P < 0.05). MiR-599 may inhibit the proliferation, invasion and promote apoptosis of bladder urothelial carcinoma J82 cells by targetingly regulating YAP1.

Keywords: Apoptosis; Invasion; Proliferation; Urothelial carcinoma of bladder; Yes-associated protein 1; microRNA-559.

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics
  • Adaptor Proteins, Signal Transducing / metabolism*
  • Apoptosis*
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / metabolism*
  • Cell Proliferation*
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • MicroRNAs / genetics*
  • Neoplasm Invasiveness
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Tumor Cells, Cultured
  • Urinary Bladder Neoplasms / genetics
  • Urinary Bladder Neoplasms / metabolism
  • Urinary Bladder Neoplasms / pathology*
  • YAP-Signaling Proteins

Substances

  • Adaptor Proteins, Signal Transducing
  • Biomarkers, Tumor
  • MIRN599 microRNA, human
  • MicroRNAs
  • Transcription Factors
  • YAP-Signaling Proteins
  • YAP1 protein, human