Evaluation of tankyrase inhibition in whole cells.
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.
Ohishi
Tomokazu
T
Division of Molecular Biotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, Tokyo, Japan.
Tsuruo
Takashi
T
Seimiya
Hiroyuki
H
D002477:Cells / Q000201:enzymology*; D018274:Electroporation; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D038501:Tankyrases / Q000037:antagonists & inhibitors*; D019098:Telomerase / Q000378:metabolism; D016615:Telomere / Q000378:metabolism
D011065:Poly(ADP-ribose) Polymerases; D038501:Tankyrases; C509186:TERT protein, human; D019098:Telomerase