Passadore M, Bianchi N, Feriotto G, Mischiati C, Rutigliano C, Gambari R
Department of Biochemistry and Molecular Biology, Ferrara, Italy.
Eur J Pharmacol. 1997 Jan 29;319(2-3):317-25. doi: 10.1016/s0014-2999(96)00849-7.
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.
在本文中，我们采用聚合酶链反应（PCR）分析了CC-1065类似物2-[[5-[(1H-吲哚-2-基]羰基)-1H-吲哚-2-基]羰基]-7-甲基-1,2,8,8a-四氢环丙[a]-吡咯并-[3,2-e]-吲哚-4-酮（U-71184）的体外序列选择性。此外，我们还确定了在从存在该药物的培养细胞中分离DNA时，是否能检测到U-71184对PCR的改变。作为分子模型系统，我们采用了人类雌激素受体基因、Ha-ras癌基因和X染色体连锁的富含（CGG）的脆性X智力低下-1基因。从我们论文中报道的实验可以得出的第一个结论是，U-71184以序列依赖性方式抑制PCR。我们实验的第二个结论是，当使用扩增雌激素受体基因区域的引物时，对来自U-71184处理细胞的DNA进行的PCR受到抑制。这种方法可能会带来关于药物在体内被靶细胞摄取以及与DNA结合的重要信息。
