Selective binding to human genomic sequences of two synthetic analogues structurally related to U-71184 and adozelesin
✍ Scribed by Nicoletta Bianchi; Giampiero Spalluto; Barbara Cacciari; Romeo Romagnoli; Giordana Feriotto; Carlo Mischiati; Cristina Rutigliano; Elena Borsetti; Pier Giovanni Baraldi; Roberto Gambari
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 390 KB
- Volume
- 46
- Category
- Article
- ISSN
- 0272-4391
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✦ Synopsis
In this paper, we analyse the in vitro sequence-selectivity of two synthetic analogues of U-71184 and adozelesin by polymerase chain reaction (PCR) performed on human genomic DNA. In addition, DNase footprinting and nucleotide sequence analysis on arrested-PCR products were performed to confirm sequence-selective binding. Finally, the antitumor effects were studied in vitro on human leukemic L1210 cells. The binding activity of the two newly synthesized compounds to human gene sequences was compared with the CC-1065 analogue U-71184, the A+T sequence-selective drug distamycin and the G+C sequence-selective drugs mithramycin and chromomycin. As molecular model systems for in vitro DNAbinding studies we used the human estrogen receptor gene and the Ha-ras oncogene. In some experiments the PCR approach was performed using as target DNA a portion of the long terminal repeat (LTR) of the human immunodeficiency type 1 virus (HIV-1). These genomic regions contain sequences that are different with respect to A+T/G+C ratios, being the upstream sequence of the human estrogen receptor gene A+T rich, while the Ha-ras and HIV-1 LTR sequences contain G+C-rich regions. The first conclusion that can be drawn from the experiments reported in our paper is that the two newly synthetized analogues of U-71184 and adozelesin inhibit PCR-mediated amplification of genomic regions in a sequence-dependent manner. A second conclusion of our experiments is that these compounds are active inhibitors of tumor cell growth in vitro.