Modulating multidrug resistance gene in leukaemia cells by short interfering RNA

Moon Nian Lim, N. S. Lau, K. M. Chang, Leong Chooi Fun, Z. Zakaria

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Introduction: The multidrug resistance gene, MDR1, is one of the genes responsible for resistance to chemotherapy in the treatment of leukaemia and other cancers. The discovery of RNA interference in mammalian cells has provided a powerful tool to inhibit the expression of this gene. However, very little is known about the transfection of leukaemia cells with short interfering RNA (siRNA) targeted at MDR1. This study aims to evaluate the effectiveness of two chemically-synthesised siRNA in modulating MDR1 gene and inhibiting P-glycoprotein expression in leukaemic cells. We also evaluated two siRNA delivery methods in this study. Methods: K562/Adr was transfected with two MDR1-targeted siRNA or negative control siRNA, by using cationic lipid-based transfection reagents or electroporator. Gene expression of MDR1 was quantified by real-time polymerase chain reaction and calculated as a percentage relative to the negative control siRNA. P-glycoprotein expression was evaluated via flow cytometry and drug sensitivity after treatment was assessed by cytotoxicity assays. Results: The percentage of MDR1 gene knockdown from cells transfected with an electroporator was significantly higher (84.4 percent, p-value is 0.094) compared to cells transfected with cationic lipid-based transfection reagents (52.8 percent). Both siRNA significantly reduced the expression of MDR1 by 84.9 percent (p-value is 0.001) and 86.0 percent (p-value is 0.011), respectively. P-glycoprotein expression was down-regulated and drug sensitivity was increased after treatment with the siRNA. Conclusion: This study shows that the two siRNA sequences are capable of modulating MDR1 and P-glycoprotein expressions and increased drug sensitivity. Transfection with an electroporator was superior to chemical transfection for leukaemia cells.

Original languageEnglish
Pages (from-to)932-938
Number of pages7
JournalSingapore Medical Journal
Volume48
Issue number10
Publication statusPublished - Oct 2007

Fingerprint

MDR Genes
Small Interfering RNA
Leukemia
Transfection
P-Glycoprotein
Pharmaceutical Preparations
Gene Knockdown Techniques
Lipids
Gene Expression
RNA Interference
Genes
Real-Time Polymerase Chain Reaction
Flow Cytometry
Therapeutics
Drug Therapy

Keywords

  • Leukaemia cells
  • Multidrug resistance gene
  • P-glycoprotein
  • Short interfering RNA transfection

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Lim, M. N., Lau, N. S., Chang, K. M., Chooi Fun, L., & Zakaria, Z. (2007). Modulating multidrug resistance gene in leukaemia cells by short interfering RNA. Singapore Medical Journal, 48(10), 932-938.

Modulating multidrug resistance gene in leukaemia cells by short interfering RNA. / Lim, Moon Nian; Lau, N. S.; Chang, K. M.; Chooi Fun, Leong; Zakaria, Z.

In: Singapore Medical Journal, Vol. 48, No. 10, 10.2007, p. 932-938.

Research output: Contribution to journalArticle

Lim, MN, Lau, NS, Chang, KM, Chooi Fun, L & Zakaria, Z 2007, 'Modulating multidrug resistance gene in leukaemia cells by short interfering RNA', Singapore Medical Journal, vol. 48, no. 10, pp. 932-938.
Lim, Moon Nian ; Lau, N. S. ; Chang, K. M. ; Chooi Fun, Leong ; Zakaria, Z. / Modulating multidrug resistance gene in leukaemia cells by short interfering RNA. In: Singapore Medical Journal. 2007 ; Vol. 48, No. 10. pp. 932-938.
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