RESUMEN
The DNA interstrand cross-links of the antitumor drug {[cis-Pt(NH3)2Cl]2(4,4'-methylenedianiline)}2+ (1) play a prevalent role in its antitumor effects. Complex 1 forms DNA long-range interstrand cross-links uniquely in the 3'-3' direction. Conformational distortions induced by these interstrand cross-links in DNA represent a potential structural motif for recognition by high-mobility-group proteins.
Asunto(s)
Aductos de ADN/química , Sustancias Intercalantes/química , Sustancias Intercalantes/farmacología , Compuestos Organoplatinos/química , Compuestos Organoplatinos/farmacología , Antineoplásicos/química , Antineoplásicos/farmacología , Secuencia de Bases , Reactivos de Enlaces Cruzados/química , Reactivos de Enlaces Cruzados/farmacología , ADN/química , Humanos , Simulación del Acoplamiento Molecular , Conformación de Ácido Nucleico/efectos de los fármacosRESUMEN
The principles of large screening strategies, which are developed by industrial companies, have been recently adopted by researchers in the fields of molecular biology and oncology as invaluable tools for translational medicine. The declining costs of laboratory robotic machines have allowed high-throughput screening to become more available to academic centres with limited resources. Here, we describe how a robotic conventional liquid handling system could be used on a daily basis in laboratories to obtain consistent and reproducible results. Our approach allowed us to quickly screen a panel of more than 20 tumorigenic and non-tumorigenic cell lines for their responses to hydroxyurea, which is a DNA-damaging anticancer therapeutic drug. The format of 384-well microplates was used for manual cell seeding, and the effect of hydroxyurea was screened at multiple concentrations. The fluorescence-based CyQuant assay was employed as the readout method to analyse the cellular DNA content. The effectiveness of our approach was demonstrated in the experimental results.