Puvvada M S, Hartley J A, Jenkins T C, Thurston D E
School of Pharmacy and Biomedical Sciences, University of Portsmouth, London, UK.
Nucleic Acids Res. 1993 Aug 11;21(16):3671-5. doi: 10.1093/nar/21.16.3671.
An assay has been developed (restriction endonuclease digestion assay--RED100) based on inhibition of the restriction endonuclease BamHI that is capable of quantitative evaluation of the relative DNA-binding affinity of pyrrolo[2,1-c] [1,4]benzodiazepine (PBD) antitumour antibiotics. This method provides comparable results to those obtained from thermal denaturation and ethidium bromide displacement assays but is much more sensitive, discriminating between molecules of similar structure such as DC-81, iso-DC-81 and neothramycin. The results reveal a trend between relative DNA-binding affinity and in vitro cytotoxicity for the PBDs in two tumour cell lines studied.
基于对限制性内切酶BamHI的抑制作用，已开发出一种检测方法（限制性内切酶消化检测法-RED100），该方法能够对吡咯并[2,1-c][1,4]苯并二氮杂卓（PBD）抗肿瘤抗生素的相对DNA结合亲和力进行定量评估。该方法所提供的结果与通过热变性和溴化乙锭置换检测法获得的结果相当，但更为灵敏，能够区分结构相似的分子，如DC-81、异-DC-81和新霉素。结果揭示了在所研究的两种肿瘤细胞系中，PBD的相对DNA结合亲和力与体外细胞毒性之间的一种趋势。
