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Altering DNA base excision repair: Use of nuclear and mitochondrial‐targeted N ‐methylpurine DNA glycosylase to sensitize astroglia to chemotherapeutic agents
Author(s) -
Harrison Jason F.,
Rinne Mikael L.,
Kelley Mark R.,
Druzhydiya M.,
Wilson Glenn L.,
Ledoux Susan P.
Publication year - 2007
Publication title -
glia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.954
H-Index - 164
eISSN - 1098-1136
pISSN - 0894-1491
DOI - 10.1002/glia.20556
Subject(s) - biology , dna repair , mitochondrion , base excision repair , apoptosis , astrocyte , dna glycosylase , programmed cell death , dna damage , microbiology and biotechnology , ap site , dna , mitochondrial dna , biochemistry , neuroscience , central nervous system , gene
Abstract Primary astrocyte cultures were used to investigate the modulation of DNA repair as a tool for sensitizing astrocytes to genotoxic agents. Base excision repair (BER) is the principal mechanism by which mammalian cells repair alkylation damage to DNA and involves the processing of relatively nontoxic DNA adducts through a series of cytotoxic intermediates during the course of restoring normal DNA integrity. An adenoviral expression system was employed to target high levels of the BER pathway initiator, N ‐methylpurine glycosylase (MPG), to either the mitochondria or nucleus of primary astrocytes to test the hypothesis that an alteration in BER results in increased alkylation sensitivity. Increasing MPG activity significantly increased BER kinetics in both the mitochondria and nuclei. Although modulating MPG activity in mitochondria appeared to have little effect on alkylation sensitivity, increased nuclear MPG activity resulted in cell death in astrocyte cultures treated with methylnitrosourea (MNU). Caspase‐3 cleavage was not detected, thus indicating that these alkylation sensitive astrocytes do not undergo a typical programmed cell death in response to MNU. Astrocytes were found to express relatively high levels of antiapoptotic Bcl‐2 and Bcl‐XL and very low levels of proapoptotic Bad and Bid suggesting that the mitochondrial pathway of apoptosis may be blocked making astrocytes less vulnerable to proapoptotic stimuli compared with other cell types. Consequently, this unique characteristic of astrocytes may be responsible, in part, for resistance of astrocytomas to chemotherapeutic agents. © 2007 Wiley‐Liss, Inc.