Specific inhibition of bcr-abl gene expression by small interfering RNA

Blood. 2003 Feb 15;101(4):1566-9. doi: 10.1182/blood-2002-06-1685. Epub 2002 Sep 26.

Abstract

Small interfering RNAs (siRNAs) were designed to target the bcr-abl oncogene, which causes chronic myeloid leukemia (CML) and bcr-abl-positive acute lymphoblastic leukemia (ALL). Chemically synthesized anti-bcr-abl siRNAs were selected using reporter gene constructs and were found to reduce bcr-abl mRNA up to 87% in bcr-abl-positive cell lines and in primary cells from CML patients. This mRNA reduction was specific for bcr-abl because c-abl and c-bcr mRNA levels remained unaffected. Furthermore, protein expression of BCR-ABL and of laminA/C was reduced by specific siRNAs up to 80% in bcr-abl-positive and normal CD34(+) cells, respectively. Finally, anti-bcr-abl siRNA inhibited BCR-ABL-dependent, but not cytokine-dependent, proliferation in a bcr-abl-positive cell line. These data demonstrate that siRNA can specifically and efficiently interfere with the expression of an oncogenic fusion gene in hematopoietic cells.

MeSH terms

  • Animals
  • Cell Division / drug effects
  • Cell Line
  • Cytokines / pharmacology
  • Fusion Proteins, bcr-abl / analysis
  • Fusion Proteins, bcr-abl / genetics*
  • Gene Expression / drug effects*
  • Green Fluorescent Proteins
  • Hematopoietic Stem Cells / chemistry
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Immunoblotting
  • Lamin Type A / genetics
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics*
  • Luminescent Proteins / genetics
  • Mice
  • Microscopy, Fluorescence
  • RNA, Messenger / analysis
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / pharmacology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Cytokines
  • Lamin Type A
  • Luminescent Proteins
  • RNA, Messenger
  • RNA, Small Interfering
  • lamin C
  • Green Fluorescent Proteins
  • Fusion Proteins, bcr-abl