Alantolactone induces cell apoptosis partially through down-regulation of testes-specific protease 50 expression

Toxicol Lett. 2014 Jan 30;224(3):349-55. doi: 10.1016/j.toxlet.2013.11.002. Epub 2013 Nov 16.

Abstract

Testes-specific protease 50 (TSP50) is aberrantly expressed in many cancer biopsies and plays a crucial role in tumorigenesis, which make it a potential cancer therapeutic target for drug discovery. Here, we constructed a firefly luciferase reporter driven by the TSP50 gene promoter to screen natural compounds capable of inhibiting the expression of TSP50. Then we identified alantolactone, a sesquiterpene lactone, could efficiently inhibit the promoter activity of TSP50 gene, further results revealed that alantolactone also efficiently inhibited the expression of TSP50 in both mRNA and protein levels. Moreover, we found alantolactone could increase the ratio of Bax/Bcl-2, and activate caspase-9 and caspase-3 in the cancer cells with high expression of TSP50, surprisingly, the same effects can also be observed in the same cells just by knockdown of TSP50 gene expression. Furthermore, our results suggested that overexpression of TSP50 decreased the cell sensitivity to alantolactone-induced apoptosis in those cancer cells. Taken together, these results suggest that alantolactone induces mitochondrial-dependent apoptosis at least partially via down-regulation of TSP50 expression.

Keywords: Alantolactone; Anticancer; Apoptosis; Drug screening; TSP50 inhibitor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antifungal Agents / toxicity*
  • Apoptosis / drug effects*
  • Blotting, Western
  • Caspase 3 / biosynthesis
  • Caspase 9 / biosynthesis
  • Cell Line, Tumor
  • Cell Membrane / drug effects
  • Cell Membrane / ultrastructure
  • Down-Regulation / drug effects
  • Humans
  • Lactones / toxicity*
  • Male
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Plasmids / genetics
  • Promoter Regions, Genetic / drug effects
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA / biosynthesis
  • RNA / isolation & purification
  • Real-Time Polymerase Chain Reaction
  • Serine Endopeptidases / biosynthesis*
  • Serine Endopeptidases / drug effects*
  • Serine Endopeptidases / genetics
  • Sesquiterpenes, Eudesmane / toxicity*
  • Transfection
  • bcl-2-Associated X Protein / genetics
  • bcl-2-Associated X Protein / metabolism

Substances

  • Antifungal Agents
  • Lactones
  • Proto-Oncogene Proteins c-bcl-2
  • Sesquiterpenes, Eudesmane
  • bcl-2-Associated X Protein
  • RNA
  • Serine Endopeptidases
  • testis-specific protease 50
  • Caspase 3
  • Caspase 9
  • alantolactone