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Selective binding to human genomic sequences of two synthetic analogues structurally related to U‐71184 and adozelesin
Author(s) -
Bianchi Nicoletta,
Spalluto Giampiero,
Cacciari Barbara,
Romagnoli Romeo,
Feriotto Giordana,
Mischiati Carlo,
Rutigliano Cristina,
Borsetti Elena,
Baraldi Pier Giovanni,
Gambari Roberto
Publication year - 1999
Publication title -
drug development research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.582
H-Index - 60
eISSN - 1098-2299
pISSN - 0272-4391
DOI - 10.1002/(sici)1098-2299(199902)46:2<96::aid-ddr2>3.0.co;2-6
Subject(s) - biology , microbiology and biotechnology , in vitro , footprinting , gene , dna , consensus sequence , retrovirus , nucleic acid sequence , genomic dna , biochemistry , genetics , peptide sequence , transcription factor
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 DNA‐binding 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. Drug Dev. Res. 46:96–106, 1999. © 1999 Wiley‐Liss, Inc.