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NLS-RARα promotes proliferation and inhibits differentiation in HL-60 cells

Xiu-Xiu Hu1, Liang Zhong2, Xi Zhang2

  • 11. Central Laboratory of Yong-chuan hospital, Chongqing Medical University, Chongqing 402160, China. ; 2. Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.

International Journal of Medical Sciences
|February 12, 2014
PubMed

Insights

Down-regulating nuclear localization signal-retinoic acid receptor alpha (NLS-RARα) inhibits leukemic cell proliferation and promotes differentiation. Conversely, over-expressing NLS-RARα enhances proliferation and hinders differentiation in acute promyelocytic leukemia models.

Area of Science:

  • Hematology
  • Molecular Biology
  • Cancer Research

Background:

  • Acute promyelocytic leukemia (APL) is characterized by a t(15;17) translocation producing a PML-RARα fusion protein.
  • Neutrophil elastase (NE) cleaves PML-RARα, yielding NLS-RARα, which localizes to the nucleus.
  • The biological role of NLS-RARα in APL pathogenesis remains largely uncharacterized.

Purpose of the Study:

  • To investigate the biological functions of the NLS-RARα fusion protein in HL-60 cells.
  • To determine the effects of NLS-RARα modulation on cell proliferation and differentiation.

Main Methods:

  • Gene silencing of NLS-RARα using short hairpin RNA (shRNA) in HL-60 cells.
  • Over-expression of NLS-RARα via recombinant adenovirus in HL-60 cells.
  • Assays included RT-PCR, Western blot, MTT assay, flow cytometry (FCM), and C-MYC expression analysis.

Main Results:

  • NLS-RARα down-regulation significantly reduced HL-60 cell proliferation and induced differentiation.
  • NLS-RARα over-expression markedly promoted HL-60 cell proliferation and inhibited ATRA-induced differentiation.
  • Over-expression of NLS-RARα led to increased C-MYC transcription and translation upon ATRA treatment.

Conclusions:

  • Down-regulation of NLS-RARα inhibits proliferation and promotes differentiation of HL-60 cells, suggesting a role in myeloid differentiation.
  • Over-expression of NLS-RARα promotes proliferation and impairs ATRA-induced differentiation, implicating it as a potential driver in APL.
  • NLS-RARα is a critical determinant of HL-60 cell behavior, influencing proliferation and differentiation pathways relevant to APL.
Keywords:
HL-60 cellNLS-RARαdifferentiationover-expressionproliferationshRNA

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