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Related Experiment Videos

Phorbol ester stimulated cathepsin L expression in U937 cells

K B Atkins1, B R Troen

  • 1Department of Internal Medicine, Veterans Administration Medical Center, University of Michigan, Ann Arbor 48109, USA.

Cell Growth & Differentiation : the Molecular Biology Journal of the American Association for Cancer Research
|June 1, 1995
PubMed
Summary

Phorbol ester (TPA) increases cathepsin L (ctsl) mRNA expression in leukemia cells via transcription. This process involves tyrosine kinase activation, independent of protein kinase C signaling.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Cathepsin L (ctsl) is a lysosomal cysteine proteinase.
  • Its synthesis and secretion are induced by transformation, growth factors, and tumor promoters.

Purpose of the Study:

  • To investigate the effect and mechanism of phorbol ester (TPA) on cathepsin L (ctsl) mRNA expression in U937 histiocytic leukemia cells.
  • To elucidate the signal transduction pathways involved in TPA-induced ctsl gene expression.

Main Methods:

  • U937 cells were treated with TPA, alone or in combination with inhibitors (cycloheximide, dexamethasone, staurosporine, H7, herbimycin A, genistein, sodium orthovanadate).
  • Cathepsin L (ctsl) mRNA expression levels were analyzed.
  • c-jun mRNA expression was also monitored.

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Main Results:

  • TPA treatment dose-dependently induced ctsl mRNA expression at the transcriptional level.
  • This induction was ablated by cycloheximide but unaffected by dexamethasone.
  • Staurosporine (a protein kinase C inhibitor) enhanced TPA-induced ctsl mRNA expression, suggesting a protein kinase C-independent pathway.
  • Tyrosine kinase inhibitors (herbimycin A, genistein) blocked TPA and staurosporine effects on ctsl mRNA.
  • Sodium orthovanadate enhanced ctsl expression induction.

Conclusions:

  • TPA-stimulated cathepsin L (ctsl) gene transcription in U937 cells is activated by a protein kinase C-independent pathway.
  • This pathway involves tyrosine kinase activation.