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Regulation of CD11b transcription by decreasing PRC2 and increased acH4 level during ATRA-induced HL-60

Huarong Tang1, Fangping Chen, Qian Tan

  • 1Department of Hematology, Xiangya Hospital, Central South University, Changsha 410008, China.

Acta Biochimica Et Biophysica Sinica
|July 7, 2009
PubMed

Insights

Polycomb repressive complex 2 (PRC2) and its H3K27me3 mark decrease during ATRA-induced HL-60 cell differentiation, correlating with increased CD11b expression and histone H4 acetylation.

Area of Science:

  • Epigenetics
  • Molecular Biology
  • Cell Differentiation

Background:

  • Polycomb repressive complex 2 (PRC2) is crucial for stem cell differentiation via histone H3 lysine 27 trimethylation (H3K27me3).
  • Understanding epigenetic regulation of gene transcription is vital for leukemia cell differentiation research.

Purpose of the Study:

  • To investigate the roles of PRC2, H3K27me3, and histone H4 acetylation (acH4) in CD11b transcription during all-trans retinoic acid (ATRA)-induced HL-60 leukemia cell differentiation.
  • To elucidate the molecular mechanisms underlying HL-60 cell reprogramming.

Main Methods:

  • Quantitative real-time polymerase chain reaction (qPCR) to measure mRNA levels.
  • Western blot analysis for protein expression.
  • Chromatin immunoprecipitation (ChIP) assay to assess protein binding and histone modifications.
  • Flow cytometry to detect cell surface marker expression.

Main Results:

  • PRC2 components (EZH2, SUZ12) mRNA and protein levels decreased significantly during ATRA treatment.
  • CD11b expression, a granulocytic marker, increased at both mRNA and protein levels.
  • ChIP assays revealed decreased SUZ12 binding and H3K27me3 at the CD11b promoter.
  • Histone H4 acetylation (acH4) levels at the CD11b promoter increased significantly.

Conclusions:

  • PRC2 activity and H3K27me3 are downregulated during ATRA-induced HL-60 differentiation.
  • Decreased H3K27me3 and increased acH4 at the CD11b promoter are associated with CD11b gene activation.
  • These histone modifications play a role in the terminal differentiation of HL-60 leukemia cells.

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