Cytotoxicity and apoptosis by survivin small interfering RNA in bladder cancer cells

BJU Int. 2010 Dec;106(11):1812-6. doi: 10.1111/j.1464-410X.2010.09259.x.

Abstract

Objective: To investigate the effects of survivin gene RNA interference on cell growth and the cell cycle in the human bladder cancer cell line T24.

Materials and methods: A small interfering RNA (siRNA) targeting survivin was transfected into T24 cells using a liposome approach. Reverse transcription-polymerase chain reaction and Western blot analysis were used to examine survivin gene expression in T24 cells. Cells densities were determined by haematocytometer counts and flow cytometry was used for cell cycle analysis. Caspase-3 activity was quantified.

Results: After treatment with survivin siRNA, the survivin gene expression in T24 cells was almost completely absent. The survivin siRNA treatment caused a profound decrease in survivin protein, which was correlated with a decrease in cell growth, G2/M arrest, and an increase in the fraction of cells undergoing apoptosis. The inhibition of survivin expression increased caspase-3 activity in T24 cells, which led to apoptosis.

Conclusions: RNA interference can efficiently suppress survivin expression in T24 cells. Targeting survivin by siRNA may be a promising approach to block proliferation of bladder cancer cells and may provide a suitable adjuvant therapy for treatment of bladder cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects*
  • Blotting, Western
  • Caspase 3 / metabolism
  • Cell Line, Tumor
  • Down-Regulation
  • Genetic Therapy*
  • Humans
  • Inhibitor of Apoptosis Proteins
  • Microtubule-Associated Proteins / genetics*
  • Microtubule-Associated Proteins / metabolism
  • RNA, Small Interfering / pharmacology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Survivin
  • Transfection
  • Urinary Bladder Neoplasms / therapy*

Substances

  • BIRC5 protein, human
  • Inhibitor of Apoptosis Proteins
  • Microtubule-Associated Proteins
  • RNA, Small Interfering
  • Survivin
  • Caspase 3