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PMID: 20028873 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Potentiation of temozolomide cytotoxicity by inhibition of DNA polymerase beta is accentuated by BRCA2 mutation.

Cancer research ·Vol. 70 ·No. 1 ·2010-01-01 ·Pages 409-17

Stachelek GC, Dalal S, Donigan KA, Campisi Hegan D, Sweasy JB, Glazer PM

Abstract

Base excision repair (BER) plays a critical role in the repair of bases damaged by oxidative metabolism or alkylating agents, such as those commonly used in cancer therapy. Incomplete BER generates intermediates that require activation of homology-dependent DNA repair to resolve. We investigated the effects of lithocholic acid (LCA), an inhibitor of the key BER enzyme DNA polymerase beta (pol beta), in cells deficient in expression of the homology-dependent repair factor BRCA2. In vitro studies show that LCA suppresses the DNA polymerase and 5'-deoxyribose phosphate lyase activities of DNA pol beta by preventing the formation of a stable pol beta-DNA complex, reducing BER effectiveness. Cytotoxicity assays based on colony formation revealed that LCA exhibits synergism with the alkylating agent temozolomide, which engages BER through DNA methylation, and that the degree of synergism is increased in cells lacking functional BRCA2. BRCA2-deficient cells also showed heightened susceptibility to both LCA and temozolomide individually. The potentiation of temozolomide cytotoxicity by LCA owes to the conversion of single-stranded DNA breaks generated through incomplete BER of methylated nucleotides into double-stranded breaks during DNA replication, as indicated by gammaH2AX immunofluorescence. Death seems to be induced in cotreated cells through an accumulation of persistent double-stranded DNA breaks. Mutations of the BRCA2 gene have been extensively characterized and are present in various cancers, implying that inhibition of BER may offer a means to augment tumor selectivity in the use of conventional cancer therapies.

MeSH Terms
Animals Antineoplastic Agents, Alkylating/administration & dosage Antineoplastic Combined Chemotherapy Protocols/pharmacology CHO Cells Cricetinae Cricetulus DNA Breaks, Double-Stranded DNA Polymerase beta/antagonists & inhibitors,drug effects DNA Repair/drug effects Dacarbazine/administration & dosage,analogs & derivatives Drug Synergism Electrophoretic Mobility Shift Assay Enzyme Inhibitors/pharmacology Fluorescent Antibody Technique Genes, BRCA2 Humans Lithocholic Acid/pharmacology Mice Mutation Temozolomide Transfection
Chemicals
Antineoplastic Agents, Alkylating Enzyme Inhibitors Lithocholic Acid Dacarbazine DNA Polymerase beta Temozolomide
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Stachelek Gregory C
Department of Therapeutic Radiology, Yale School of Medicine, New Haven, Connecticut 06520-8040, USA.
Dalal Shibani
Donigan Katherine A
Campisi Hegan Denise
Sweasy Joann B
Glazer Peter M
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Article Info
Journal
Cancer research
Abbr.
Cancer Res
ISSN
1538-7445
DOI
10.1158/0008-5472.CAN-09-1353
Published
2010-01-01
Epub
2009-00-22
Pages
409-17
Language
English
Region
United States
NLM ID
2984705R
PMCID
PMC2943728
Subset
IM
Grants
NIEHS NIH HHS · R01 ES005775 · United States
NCI NIH HHS · P01CA129186 · United States
NCI NIH HHS · P01 CA129186 · United States
NIGMS NIH HHS · T32 GM007205 · United States
NCI NIH HHS · P01 CA129186-03 · United States
NIEHS NIH HHS · R01 ES005775-17 · United States
NIEHS NIH HHS · R01ES05775 · United States
External Links
PubMed source DOI: 10.1158/0008-5472.CAN-09-1353 PMC Full Text View journal details
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