Bcl-2 targeting siRNA expressed by a T7 vector system inhibits human tumor cell growth in vitro

Int J Oncol. 2004 Mar;24(3):615-21.

Abstract

A T7 promoter driven siRNA expression vector system (Bcl-2/T7) that targets Bcl-2 mRNA in MCF-7 human cancer cells was designed in the present study. In the presence of prebound T7 RNA polymerase, successful expression of Bcl-2 siRNA as well as its function was demonstrated via cell proliferation assays, Bcl-2 Elisa, and TUNEL assay. MCF-7 breast cancer cells transfected with Bcl-2/T7 show decreased levels of Bcl-2 expression at the protein level as well as decreased cell proliferation. Also, the number of apoptotic cells was increased in cells expressing Bcl-2 siRNA. Previous studies have shown that Bcl-2 levels are increased in a large number of different types of cancer. Therefore, the ability of Bcl-2/T7 to produce functional Bcl-2 siRNA in breast cancer cells suggests a potential role for this delivery system in cancer gene therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis
  • Base Sequence
  • Breast Neoplasms / therapy*
  • Cell Division
  • Cell Line, Tumor
  • Cell Separation
  • DNA-Directed RNA Polymerases / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Genetic Therapy / methods*
  • Green Fluorescent Proteins
  • HeLa Cells
  • Humans
  • In Situ Nick-End Labeling
  • Luminescent Proteins / metabolism
  • Molecular Sequence Data
  • Plasmids / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / biosynthesis*
  • Proto-Oncogene Proteins c-bcl-2 / genetics*
  • RNA, Small Interfering / metabolism*
  • Time Factors
  • Transcription, Genetic
  • Viral Proteins

Substances

  • Luminescent Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Small Interfering
  • Viral Proteins
  • Green Fluorescent Proteins
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases