In vitro and in vivo binding of a CC-1065 analogue to human gene sequences: a polymerase-chain reaction study.
In this paper we analyse the in vitro sequence selectivity of the CC-1065 analogue 2-[[5-[(1H-indol-2-yl]carbonyl)-1H-indol-2-yl] carbonyl]-7-methyl-1,2,8,8a-tetrahydrocyclopropa [c]-pyrrolo-[3,2-e]-indol-4-one (U-71184) employing 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 systems we employed the human estrogen receptor gene, the Ha-ras oncogene and the chromosome X-linked, (CGG)-rich fragile X mental retardation-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-dependent manner. A second conclusion of our experiments is that PCR performed on DNA from U-71184-treated cells is inhibited when the primers amplifying the estrogen receptor gene region are used. This approach might bring important information on both in vivo uptake of the drug by target cells and binding to DNA.
Passadore
M
M
Department of Biochemistry and Molecular Biology, Ferrara, Italy.
Bianchi
N
N
Feriotto
G
G
Mischiati
C
C
Rutigliano
C
C
Gambari
R
R
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000494:pharmacology; D001483:Base Sequence; D001943:Breast Neoplasms / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D004214:Distamycins / Q000494:pharmacology; D000080890:Duocarmycins; D005600:Fragile X Syndrome / Q000235:genetics; D011905:Genes, ras / Q000235:genetics; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D007933:Leucomycins / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011960:Receptors, Estrogen / Q000235:genetics
D000900:Anti-Bacterial Agents; D000903:Antibiotics, Antineoplastic; D000970:Antineoplastic Agents; D004214:Distamycins; D000080890:Duocarmycins; D007211:Indoles; D007933:Leucomycins; D011960:Receptors, Estrogen; C054483:U 71184; C018782:CC 1065; D004247:DNA