Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity

World J Gastroenterol. 2006 May 14;12(18):2901-7. doi: 10.3748/wjg.v12.i18.2901.

Abstract

Aim: To investigate the inhibitory effect of small interfering RNA (siRNA) on the expression of survivin in pancreatic cancer cell line PC-2 and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity.

Methods: A siRNA plasmid expression vector against survivin was constructed and transfected into PC-2 cells with Lipofectamine 2000. The down regulation of survivin expression was detected by semi-quantitative RT-PCR and immunohistochemical SP method and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity was detected by flow cytometry.

Results: The sequence-specific siRNA efficiently and specifically down-regulated the expression of survivin at both mRNA and protein levels. The expression inhibition ratio was 81.25% at mRNA level detected by semi-quantitative RT-PCR and 74.24% at protein level detected by immunohistochemical method. Forty-eight hours after transfection,apoptosis was induced in 7.03% cells by siRNA and in 14.58% cells by siRNA combined with radiation.

Conclusion: The siRNA plasmid expression vector against survivin can inhibit the expression of survivin in PC-2 cells efficiently and specifically. Inhibiting the expression of survivin can induce apoptosis of PC-2 cells and enhance its radiosensitivity significantly. RNAi against survivin is of potential value in gene therapy of pancreatic cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / physiology*
  • Cell Line, Tumor
  • Down-Regulation
  • Flow Cytometry
  • Gene Expression Regulation, Neoplastic / drug effects*
  • Gene Expression Regulation, Neoplastic / physiology
  • Genetic Therapy
  • Genetic Vectors
  • Humans
  • Immunohistochemistry
  • Inhibitor of Apoptosis Proteins
  • Microtubule-Associated Proteins / analysis
  • Microtubule-Associated Proteins / genetics*
  • Microtubule-Associated Proteins / physiology
  • Neoplasm Proteins / analysis
  • Neoplasm Proteins / genetics*
  • Neoplasm Proteins / physiology
  • Pancreatic Neoplasms / chemistry
  • Pancreatic Neoplasms / pathology*
  • Pancreatic Neoplasms / physiopathology
  • Pancreatic Neoplasms / radiotherapy
  • Plasmids
  • RNA, Messenger / analysis
  • RNA, Small Interfering / pharmacology*
  • Radiation Tolerance / physiology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Survivin
  • Transfection

Substances

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