Morpholino Oligo Antisense efficiently suppresses BCR/ABL and cell proliferation in CML: specific inhibition of BCR-ABL gene expression by Morpholino Oligo Antisense in BCR-ABL(+) cells

Hematology. 2012 Jan;17(1):28-34. doi: 10.1179/102453311X13127324303470.

Abstract

Chronic myeloid leukemia is a disorder that develops when a hematopoietic stem cell acquires the Philadelphia chromosome carrying the chimeric BCR/ABL oncogene leading to a deregulated cell proliferation and a decreased apoptosis in response to mutagenic stimuli. Therefore, it has been considered that BCR/ABL oncogene is a potential attractive target for anticancer agents. Antisense strategies aiming to suppress the expression of BCR/ABL in chronic myeloid leukemia cells have been studied by several research groups over the last decade. In the present study, the effect of Morpholino Oligo Antisense in BCR/ABL oncogene silencing was evaluated. To examine the hypothesis, K562 was used as a BCR/ABL fusion gene positive cell line using a Jurkat cell line as a control. The capacity of Morpholino Oligo Antisense in inhibiting the translation of p210(bcr-abl) protein by a western blotting technique, inhibition of cell proliferation, and stimulation of apoptosis by flow cytometric analysis after 24 and 48 hours was studied. Prolonged exposure of K562 cell line to Morpholino Oligo Antisense targeted against BCR-ABL showed proliferation inhibition as the main feature. Following western blotting, we found that complete silencing of BCR-ABL had been achieved but flow cytometric analysis showed no significant apoptosis. The results indicate that Morpholino Oligo Antisense was able to inhibit p210(bcr-abl), but did not induce apoptosis due to co-silencing of BCR.

MeSH terms

  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Base Sequence
  • Cell Growth Processes / drug effects
  • Cell Growth Processes / genetics
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • Flow Cytometry
  • Fusion Proteins, bcr-abl / antagonists & inhibitors*
  • Fusion Proteins, bcr-abl / biosynthesis
  • Fusion Proteins, bcr-abl / genetics*
  • Gene Expression
  • Humans
  • Jurkat Cells
  • K562 Cells
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics*
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / metabolism
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / pathology
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / therapy*
  • Morpholinos / genetics
  • Morpholinos / pharmacology*
  • Oligonucleotides, Antisense / genetics
  • Oligonucleotides, Antisense / pharmacology*

Substances

  • Morpholinos
  • Oligonucleotides, Antisense
  • Fusion Proteins, bcr-abl