IN-VITRO AND IN-VIVO BINDING OF A CC-1065 ANALOG TO HUMAN GENE-SEQUENCES - A POLYMERASE-CHAIN REACTION STUDY

In this paper we analyse the in vitro sequence selectivity of the CC-1 065 analogue 2-[[5-[(1H-indol-2-ylcarbonyl)-1H-indol-2 7-methy1-1,2,8, 8a-tetrahydrocyclopropa[c]-pyrrolo- [3,2-e]-indol-4-one (U-71184) empl oying the polymerase-chain reaction (PCR). In addition, we determined whether alteration of PCR by U-71184 is detected when DNA is isolated from cells cultured in the presence of this drug. As molecular model s ystems we employed the human estrogen receptor gene, the Ha-ras oncoge ne and the chromosome X-linked, (CGG)-rich fragile X mental retardatio n-1 gene. The first conclusion that can be drawn from the experiments reported in our paper is that U-71184 inhibits PCR in a sequence-depen dent manner. A second conclusion of our experiments is that PCR perfor med on DNA from U-71184-treated cells is inhibited when the primers am plifying the estrogen receptor gene region are used. This approach mig ht bring important information on both in vivo uptake of the drug by t arget cells and binding to DNA.