Preferential binding of cisplatin to mitochondrial DNA of Chinese hamster ovary cells.
Some chemical carcinogens localize preferentially in mitochondrial DNA (mtDNA) when compared with genomic DNA (gDNA). Here we compare the ability of cisplatin (cis-diamminedichloroplatinum[II]) to induce DNA adducts in both genomic and mtDNA of Chinese hamster ovary (CHO) cells in culture. Cytotoxicity was examined by cell survival 4, 8 and 24 h after exposure to 50 microM cisplatin. Cisplatin-DNA adducts were measured in DNA from nuclear and mitochondrial fractions by dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA), a sensitive competitive microtiter-based immunoassay utilizing antiserum elicited against cisplatin-modified DNA. An additional comparison of cisplatin-DNA binding in both compartments was performed by immunoelectron microscopy using the cisplatin-DNA antiserum and colloidal gold. DELFIA analysis of cisplatin-DNA adducts in gDNA and mtDNA showed a six-fold higher incorporation of drug into mtDNA as compared to gDNA. Morphometric studies of colloidal gold distribution in photomicrographs of CHO cells showed mtDNA to contain a four-fold higher concentration of cisplatin as compared to nuclear DNA. Therefore, both methods demonstrated a preferential binding of cisplatin to mtDNA versus gDNA.
Olivero
O A
OA
Laboratory of Cellular Carcinogenesis and Tumor Promotion, NIH, National Cancer Institute, Bethesda, MD 20892, USA.
Semino
C
C
Kassim
A
A
Lopez-Larraza
D M
DM
Poirier
M C
MC
D000818:Animals; D016466:CHO Cells; D002467:Cell Nucleus / Q000235:genetics / Q000648:ultrastructure; D002945:Cisplatin / Q000378:metabolism* / Q000633:toxicity; D006224:Cricetinae; D018736:DNA Adducts; D004272:DNA, Mitochondrial / Q000378:metabolism*; D015200:Fluoroimmunoassay; D016678:Genome; D016253:Microscopy, Immunoelectron; D008928:Mitochondria / Q000235:genetics / Q000648:ultrastructure
D018736:DNA Adducts; D004272:DNA, Mitochondrial; D002945:Cisplatin