Evaluation of tankyrase inhibition in whole cells

Methods Mol Biol. 2007:405:133-46. doi: 10.1007/978-1-60327-070-0_11.

Abstract

The telomeric poly(ADP-ribose) polymerase (PARP), tankyrase 1, modulates the impact of telomerase inhibition on human cancer cells. Thus, overexpression of tankyrase 1 in telomerase-positive cancer cells confers resistance to telomerase inhibitors, such as MST-312, whereas pharmacological inhibition of tankyrase 1 enhances telomere shortening by MST-312. These facts indicate that tankyrase 1 could be a target for telomere-directed molecular cancer therapy. Here, the authors describe a convenient method to monitor the telomeric function of tankyrase 1. This protocol takes much less time than the telomere Southern blot analysis and can be utilized as a rapid screening system for tankyrase 1 inhibitors that are effective in intact cells. For direct monitoring of tankyrase 1 PARP activity, a protocol for the in vitro enzyme assay is also described.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells / enzymology*
  • Electroporation
  • Fluorescent Antibody Technique
  • HeLa Cells
  • Humans
  • Molecular Biology / methods*
  • Poly(ADP-ribose) Polymerases / metabolism
  • Tankyrases / antagonists & inhibitors*
  • Telomerase / metabolism
  • Telomere / metabolism

Substances

  • Poly(ADP-ribose) Polymerases
  • Tankyrases
  • TERT protein, human
  • Telomerase