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Retinoid receptor antisense DNAs inhibit alkaline phosphatase induction and clonogenicity in malignant keratinocytes

F O Cope1, J J Wille

  • 1Southern Research Institute, Birmingham, AL 35255-5305.

Proceedings of the National Academy of Sciences of the United States of America
|July 1, 1989
PubMed

Insights

Antisense oligodeoxynucleotides targeting cellular retinol-binding protein I (cRBP) and nuclear retinoic acid receptor alpha (hnRAR) inhibited retinoid signaling in malignant keratinocytes. This disruption affected cell growth, differentiation, and clonogenic potential, highlighting the receptors' crucial roles.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Dermatology

Background:

  • Retinoids, vitamin A derivatives, play critical roles in cellular processes.
  • Cellular retinol-binding protein I (cRBP) and nuclear retinoic acid receptor alpha (hnRAR) are key mediators of retinoid signaling.
  • Understanding these receptors' functions is vital for cellular growth and differentiation research.

Purpose of the Study:

  • To investigate the role of cRBP and hnRAR in retinoid signal transduction.
  • To determine the effects of inhibiting cRBP and hnRAR on malignant keratinocyte behavior.
  • To explore the potential of antisense oligodeoxynucleotides as tools to study retinoid receptor function.

Main Methods:

  • Synthesis of antisense and sense oligodeoxynucleotides targeting cRBP and hnRAR.
  • Treatment of human malignant keratinocytes with synthesized oligodeoxynucleotides.
  • Assessment of cRBP and hnRAR levels via Western blotting or similar techniques.
  • Evaluation of cellular responses including alkaline phosphatase induction, cell morphology changes, and clonogenic potential.

Main Results:

  • Antisense oligodeoxynucleotides significantly reduced cytoplasmic cRBP and hnRAR levels in a dose- and time-dependent manner.
  • Retinol-induced alkaline phosphatase activity was blocked by antisense treatment, but not by sense oligodeoxynucleotides.
  • Antisense treatment induced morphological changes like cell rounding and loss of adhesion, and inhibited clonogenic potential.
  • Control treatments with cytochalasin B or sodium azide did not affect cRBP or hnRAR levels despite causing morphological changes.

Conclusions:

  • Both cytoplasmic and nuclear retinoid receptors are essential for mediating retinoid responses critical for keratinocyte growth and differentiation.
  • Targeting cRBP and hnRAR with antisense oligodeoxynucleotides offers a method to probe retinoid signaling pathways.
  • Specific gene regulation by retinoids and their receptors likely underlies the diverse effects of vitamin A compounds.

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