Formation and Repair of Antitumor Antibiotic CC-1065-Induced DNA Adducts in the Adenine Phosphoribosyltransferase and Amplified Dihydrofolate Reductase Genes of Chinese Hamster Ovary Cells

• DOI: 10.1021/bi00175a048
• Publication Date: 2005-03-16T19:59:31Z
• Authors: Moon Shong Tang; Ming Qian; Annie Pao
• Keywords: 0303 health sciences; Antibiotics, Antineoplastic; Indoles; DNA Repair; Dose-Response Relationship, Drug; Adenine Phosphoribosyltransferase; Gene Amplification; CHO Cells; Leucomycins; 3. Good health; Duocarmycins; Tetrahydrofolate Dehydrogenase; 03 medical and health sciences; Cricetinae; Animals; DNA Damage

CC-1065 is a potent antitumor antibiotic which bonds to duplex DNA specifically; the biological effects of the drug are presumably the consequences of its DNA interactions. In order to investigate the factors which may affect drug-DNA bonding in cells, a method using a thermal-alkaline treatment to induce phosphodiester bond breakage at the drug-DNA bonding sites and Southern DNA transfer-hybridization to quantify drug-DNA bonding at defined sequences in drug-treated cultured mammalian cells was developed. We have found that in vivo, in cultured Chinese hamster ovary (CHO) cells, CC-1065 bonds twice as efficiently in the highly amplified dihydrofolate reductase (DHFR) gene domains as in the nonamplified adenine phosphoribosyltransferase (APRT) gene domain. However, in vitro, in purified CHO cellular DNA, CC-1065 bonds equally to both the DHFR and APRT genes. We observed a significant degree of "gene-specific" preferential repair for drug-DNA adducts in the amplified DHFR gene domains, and it appears that this "gene-specific" repair reflects "transcribed-strand specific" repair. These results suggest that DNA amplification may affect drug-DNA adduct formation and transcription may affect its repair.