Effects of siRNAs in combination with Gleevec on K-562 cell proliferation and Bcr-Abl expression

Benjamin Baker, Daniel Kestler, Albert T. Ichiki

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

RNA interference (RNAi) is the repression of gene expression through a cellular mechanism of transcript-specific mRNA degradation. RNAi has been observed in human cells and applied to the modulation of a variety of human transcripts. Our goals were to deliver small interfering RNA (siRNA) using a liposome-based method, and to show Bcr-Abl siRNA specificity against K-562 cells, alone or in combination with Gleevec. Both synthetic (syn) siRNA, consisting of homogeneous 21-nucleotide-long RNA duplexes specific for the Bcr-Abl fusion site, and recombinant (r)-generated Bcr-Abl siRNA were employed. siRNA was transfected into K-562 cells with greater than 90% efficiency using RNAiFect™, as judged by fluorescence analysis. The Bcr-Abl transcript was inhibited using either siRNA preparation as measured by RT-PCR or real-time PCR. The IC50 of Gleevec in the K-562 subline F1 was lowered over 3-fold from 0.2 to 0.06 μM in cells transfected with either syn or rBcr-Abl siRNA. No effect was observed in cells after transfection with an irrelevant control siRNA. Therefore, K-562 cells transfected with RNAifect deliver Bcr-Abl siRNA efficiently and the Bcr-Abl siRNA decreased the IC 50 of Gleevec required to inhibit the high levels of Bcr-Abl protein found in K-562 cells.

Original languageEnglish (US)
Pages (from-to)499-507
Number of pages9
JournalJournal of Biomedical Science
Volume13
Issue number4
DOIs
StatePublished - Jul 1 2006

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Cell proliferation
Small Interfering RNA
Cell Proliferation
RNA
RNA Interference
Imatinib Mesylate
RNA Stability
Gene expression
Liposomes
Inhibitory Concentration 50
Transfection
Real-Time Polymerase Chain Reaction
Fusion reactions
Nucleotides
Fluorescence
Cells
Modulation
Gene Expression
Degradation
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Effects of siRNAs in combination with Gleevec on K-562 cell proliferation and Bcr-Abl expression. / Baker, Benjamin; Kestler, Daniel; Ichiki, Albert T.

In: Journal of Biomedical Science, Vol. 13, No. 4, 01.07.2006, p. 499-507.

Research output: Contribution to journalArticle

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