SiRNA-mediated silencing of Snail-1 induces apoptosis and alters micro RNA expression in human urinary bladder cancer cell line

Artif Cells Nanomed Biotechnol. 2017 Aug;45(5):969-974. doi: 10.1080/21691401.2016.1198361. Epub 2016 Jun 20.

Abstract

Snail-1 known as one of the important transcription factor is a mediator of survival and cell migration, and expression is raised in numerous cancer types. Snail-1 gene may show a role in recurrence of several cancers including bladder cancer by down-regulating E-cadherin, inducing an epithelial to mesenchymal transition (EMT) and its related microRNAs (miRNAs). The aim of this study was to investigate the effect of a specific Snail-1 siRNA on apoptosis and alter EMT related miRNAs of EJ-138 (bladder cancer) cells. The cells were transfected with siRNAs using transfection reagent. The cytotoxic effects of Snail-1 siRNA, on bladder cancer cells were determined using MTT assay. Relative Snail-1 mRNA levels were measured by QRT- PCR, respectively. Apoptosis was measured by TUNEL test based on labeling of DNA strand breaks. We also evaluated miR-29b, miR-21, and miR-203 expression by QRT-PCR to determine alteration in miRNAs expression involved in EMT. Snail-1 siRNA significantly reduced mRNA expression levels in 48 h after transfection at the concentration of 60 pmol in bladder cancer cells. We also showed that the silencing of Snail-1 led to the induction of apoptosis. miR-21 and miR-29b depression have been shown in Snail-1 suppressed group in EJ-138 cells in vitro. These results propose that Snail-1 might play an important role in the progression of bladder cancer, and be a potential therapeutic target for trigger apoptosis and suppression of EMT-related miRNAs in bladder cancer.

Keywords: Apoptosis; EMT; bladder cancer; micro RNA; small interference RNA; snail-1.

MeSH terms

  • Apoptosis / genetics*
  • Cadherins / genetics
  • Cell Line, Tumor
  • Down-Regulation / genetics
  • Gene Expression Regulation, Neoplastic / genetics*
  • Gene Silencing*
  • Humans
  • MicroRNAs / genetics*
  • Neoplasm Invasiveness
  • RNA, Small Interfering / genetics*
  • Snail Family Transcription Factors / deficiency
  • Snail Family Transcription Factors / genetics*
  • Urinary Bladder Neoplasms / pathology*

Substances

  • Cadherins
  • MIRN21 microRNA, human
  • MIRN29a microRNA, human
  • MicroRNAs
  • RNA, Small Interfering
  • SNAI1 protein, human
  • Snail Family Transcription Factors