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Regulation and expression of the ATP-binding cassette transporter ABCG2 in human embryonic stem cells

Raji Padmanabhan1, Kevin G Chen, Jean-Pierre Gillet

  • 1Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.

Stem Cells (Dayton, Ohio)
|August 14, 2012
PubMed

Insights

Human embryonic stem cells (hESCs) express ABCG2 mRNA but not protein, which is regulated by specific microRNAs during differentiation. This clarifies ABCG2 expression and reveals miRNA control over transporter mRNA translation in hESCs.

Area of Science:

  • Stem cell biology
  • Molecular and cellular biology
  • Biochemistry

Background:

  • Multidrug transporters like ABCG2 are crucial in various cell types.
  • ABCG2 expression in human embryonic stem cells (hESCs) is poorly understood, with conflicting prior research.
  • Clarifying ABCG2 in hESCs is vital for understanding stem cell function and drug resistance.

Purpose of the Study:

  • To definitively investigate the expression and regulation of ABCG2 in human embryonic stem cells (hESCs).
  • To resolve conflicting literature data on ABCG2 presence and function in hESCs.
  • To explore the role of microRNAs in controlling ABCG2 translation in hESCs.

Main Methods:

  • Quantitative real-time PCR and Northern blotting for mRNA detection.
  • Whole genome RNA sequencing for comprehensive transcript analysis.
  • Western blotting and immunofluorescence microscopy for protein localization.
  • MicroRNA mimic and inhibitor transfections to assess regulatory roles.

Main Results:

  • Full-length ABCG2 mRNA was detected in undifferentiated hESCs.
  • ABCG2 protein remained undetectable, even with differentiation or drug treatment.
  • Surface ABCG2 protein appeared during differentiation, correlating with Stage-Specific Embryonic Antigen-1 (SSEA-1) expression.
  • This protein expression was linked to the downregulation of hsa-miR-519c and hsa-miR-520h.
  • miRNA manipulation confirmed their direct role in regulating ABCG2 translation.

Conclusions:

  • ABCG2 expression in hESCs is primarily at the mRNA level, with protein translation being tightly controlled.
  • ABCG2 protein expression emerges during differentiation, influenced by BMP-4 signaling and miRNA activity.
  • Specific miRNAs (hsa-miR-519c, hsa-miR-520h) directly inhibit ABCG2 translation in hESCs.
  • This study provides novel insights into miRNA-mediated translational control of membrane transporters in human embryonic stem cells.

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