DNA repair: counteragent in mutagenesis and carcinogenesis-- accomplice in cancer therapy resistance.
DNA-reactive carcinogens and anticancer drugs induce many structurally distinct mutagenic and cytotoxic DNA lesions. The varying capability of normal and malignant cells to recognize and repair specific DNA lesions influences both cancer risk and the relative sensitivity or resistance of cancer cells towards cytotoxic agents. Using monoclonal antibody-based immunoanalytical assays, very low amounts of defined carcinogen-DNA adducts can be quantified in bulk genomic DNA, in individual genes, and in the nuclear DNA of single cells. DNA repair kinetics can, thus, be measured in a lesion-, gene-, and cell type-specific manner, and the DNA repair profiles of malignant cells can be monitored in individual patients. Even structurally very similar DNA lesions may be repaired with strongly differing efficiency. The miscoding DNA alkylation products O(6)-methylguanine and O(6)-ethylguanine, for example, differ only by one CH(2) group. These lesions are formed in DNA upon exposure to N-methyl-N-nitrosourea or N-ethyl-N-nitrosourea, both of which induce mammary adenocarcinomas in female rats at high yield. Unrepaired O(6)-alkylguanines in DNA cause G:C-->A:T transition mutations via mispairing during DNA replication. O(6)-methylguanines are repaired at a similar slow rate in both transcriptionally active (H-ras, beta-actin) and inactive genes (IgE heavy chain; bulk DNA) of the target mammary epithelia (which express the repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT) at a very low level). In contrast, O(6)-ethylguanines are repaired approximately 20 times faster than O(6)-methylguanines in both DNA strands of the transcribed genes selectively via an AGT-independent, as yet unclarified excision mechanism. Accordingly, G:C-->A:T transitions resulting from the misreplication of an O(6)-methylated guanine at the second position of codon 12 (GGA) of H-ras represent a frequent "signature" mutation in rat mammary adenocarcinomas that develop after exposure to N-methyl-N-nitrosourea. However, this mutation is not observed when these tumors are induced by N-ethyl-N-nitrosourea, due to the fast repair of O(6)-ethylguanines in the H-ras gene. The key importance of "conventional" and "conditional" gene knockout technology for resolving the intricacies of the complex network of DNA repair pathways is briefly discussed.
Rajewsky
M F
MF
Institute of Cell Biology (Cancer Research) [IFZ], University of Essen Medical School and West German Cancer Center Essen, Hufeland-Strasse 55, D-45122, Essen, Germany. rajewsky@uni-essen.de
Engelbergs
J
J
Thomale
J
J
Schweer
T
T
D000230:Adenocarcinoma / Q000139:chemically induced / Q000378:metabolism; D000477:Alkylating Agents / Q000008:administration & dosage / Q000633:toxicity; D000818:Animals; D002273:Carcinogens / Q000008:administration & dosage / Q000633:toxicity; D004247:DNA / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D018736:DNA Adducts / Q000737:chemistry / Q000378:metabolism; D004260:DNA Repair; D019008:Drug Resistance, Neoplasm; D005038:Ethylnitrosourea / Q000009:adverse effects / Q000633:toxicity; D005260:Female; D011905:Genes, ras / Q000187:drug effects / Q000235:genetics; D006147:Guanine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008325:Mammary Neoplasms, Experimental / Q000139:chemically induced / Q000378:metabolism; D008770:Methylnitrosourea / Q000008:administration & dosage / Q000633:toxicity; D009153:Mutagens / Q000008:administration & dosage / Q000633:toxicity; D009154:Mutation; D051381:Rats
D000477:Alkylating Agents; D002273:Carcinogens; D018736:DNA Adducts; D009153:Mutagens; C007673:6-ethylguanine; D006147:Guanine; D008770:Methylnitrosourea; D004247:DNA; C008449:O-(6)-methylguanine; D005038:Ethylnitrosourea