Reduced DNA double strand breaks in chlorambucil resistant cells are related to high DNA-PKcs activity and low oxidative stress》.pdf

Toxicology 193 (2003) 137–152 Reduced DNA double strand breaks in chlorambucil resistant cells are related to high DNA-PKcs activity and low oxidative stress Istvan Boldogh a,∗, Gargi Roy c , Myung-Soog Lee a , Attila Bacsi d , Tapas K. Hazrab , Kishor K. Bhakat b , Gokul C. Das a , Sankar Mitrab a Department of Microbiology and Immunology, Sealy Center for Molecular Sciences, University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA b Department of Human Biological Chemistry and Genetics, Sealy Center for Molecular Sciences, University of Texas Medical Branch at Galveston, Galveston, TX 77555, USA c Acorda Therapeutics Incorporated, Hawthorne, NY 10532, USA d School of Medicine at Debrecen, Debrecen 4012, Hungary Abstract Modulation of DNA repair represents a strategy to overcome acquired drug resistance of cells to genotoxic chemotherapeutic agents, including nitrogen mustards (NM). These agents induce DNA inter-strand cross-links, which in turn produce double strand breaks (dsbs). These breaks are primarily repaired via the nonhomologous end-joining (NHEJ) pathway. A DNA-dependent protein kinase (DNA-PK) complex plays an important role in NHEJ, and its increased level/activity is associated with acquired drug resistance of human tumors. We show in this report that the DNA-PK complex has comparable levels and kinase activity of DNA-PK catalytic subunit (DNA-PKcs) in a nearly isogenic pair of drug-sensitive (A2780) and resistant (A2780/100) cells; however, treatment with chlorambucil (Cbl), a NM-type of drug, induced differential effects in these cells. The kinase activity of DNA-PKcs was increased up to 2 h after Cbl treatment in both cell types; however, it