Preferential repair of the N-ras gene in K 562 cells after exposure to cisplatin.
Cisplatin is one of the most active and widely used anticancer drugs; however, its clinical efficiacy is often limited by the development of resistance. Since several studies indicated that ras oncogenes may modulate the cellular response to cisplatin or radiation, we investigated the gene-specific repair of the N-ras gene in the human K 562 cell line after exposure to cisplatin using a novel nonradioactive polymerase chain reaction inhibition assay. This assay is based on the fact that DNA lesions can block the Taq polymerase and thereby result in a decreased amplification of a damaged segment compared to the amplification of the same segment in a non-damaged template. The overall genomic repair rate was measured by atomic absorption spectroscopy. Immediately after cisplatin exposure no amplification segment was observed. However, a complete restoration of the N-ras gene (2.4 kb) was seen after 8 h posttreatment incubation. In contrast, only 60% of the overall genome was repaired at this time. Our results clearly indicate that cisplatin-induced DNA lesions are more efficiently removed from transcribed regions within the DNA, suggesting that the efficiency of DNA repair in a given gene may be correlated to its transcriptional activity. Since ras oncogenes control several cellular signal transduction pathways, known to be involved in DNA damage response, preferential repair of the N-ras gene could therefore be an important step to prevent inactivation of cellular defense mechanisms after exposure to genotoxic agents.
Dempke
W
W
Department of Hematology and Oncology, Martin Luther University, Halle/Saale, Germany.
Voigt
W
W
Grothey
A
A
Schmoll
H J
HJ
D000970:Antineoplastic Agents / Q000494:pharmacology*; D016375:Antisense Elements (Genetics); D015139:Blotting, Southern; D002945:Cisplatin / Q000494:pharmacology*; D017931:DNA Primers; D004260:DNA Repair / Q000187:drug effects*; D004273:DNA, Neoplasm / Q000187:drug effects*; D011905:Genes, ras / Q000187:drug effects*; D006801:Humans; D020014:K562 Cells; D007951:Leukemia, Myeloid / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000970:Antineoplastic Agents; D016375:Antisense Elements (Genetics); D017931:DNA Primers; D004273:DNA, Neoplasm; D002945:Cisplatin