Measurement of Drug-Stabilized Topoisomerase II Cleavage Complexes by Flow Cytometry.
Curr Protoc Cytom
; 81: 7.48.1-7.48.8, 2017 Jul 05.
Article
in En
| MEDLINE
| ID: mdl-28678420
ABSTRACT
The poisoning of Topoisomerase II (Top2) has been found to be useful as a therapeutic strategy for the treatment of several tumors. The mechanism of Top2 poisons involves a drug-mediated stabilization of a Top2-DNA complex, termed Top2 cleavage complex (Top2cc), which maintains a 5' end of DNA covalently bound to a tyrosine from Top2 through a phosphodiester group. Drug-stabilized Top2cc leads to Top2-linked-DNA breaks, which are believed to mediate their cytotoxicity. Several time-consuming or cell type-limiting assays have been used in the past to study drug-stabilized Top2cc. Here, we describe a flow cytometry-based method that allows a rapid assessment of drug-induced Top2cc, which is suitable for high throughput analysis in almost any kind of human cell. The analyses of the drug-induced Top2cc in the cell cycle context and the possibility to track its removal are additional benefits from this methodology. © 2017 by John Wiley & Sons, Inc.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
DNA
/
DNA Topoisomerases, Type II
/
Etoposide
/
Flow Cytometry
Limits:
Animals
/
Humans
Language:
En
Year:
2017
Type:
Article