Fujiwara Y, Kasahara K, Sugimoto Y, Nishio K, Ohmori T, Saijo N
Pharmacology Division, National Cancer Center Research Institute, Tokyo.
Jpn J Cancer Res. 1990 Dec;81(12):1210-3. doi: 10.1111/j.1349-7006.1990.tb02680.x.
Using a gel mobility shift assay, we have identified proteins, in the nuclear extracts of a human lung cancer cell line, that recognize cis-diamminedichloroplatinum(II) (cis-DDP, CDDP)-modified DNA. A 158-base-pair double-stranded DNA fragment, derived from pBR322 plasmid DNA, was modified by either CDDP, tetrachloro(dl-trans)-1,2-diaminocyclohexaneplatinum(IV) (tetraplatin) or trans-DDP (the stereoisomer of CDDP and clinically ineffective). These platinum drug-modified probes were incubated with nuclear extracts and analyzed by gel mobility shift assay. Proteins in the extracts selectively recognized the clinically active platinum-modified DNA fragment. No binding to the trans-DDP-modified DNA fragment was observed. These proteins may play a role in the cytotoxicity or in a DNA repair process.
利用凝胶迁移率变动分析，我们在一种人肺癌细胞系的核提取物中鉴定出了能识别顺二氯二氨铂（II）（顺铂，CDDP）修饰DNA的蛋白质。一段源自pBR322质粒DNA的158碱基对双链DNA片段，用顺铂、四氯（反式）-1,2-二氨基环己烷铂（IV）（四铂）或反式顺铂（顺铂的立体异构体且临床无效）进行修饰。这些铂类药物修饰的探针与核提取物一起孵育，并通过凝胶迁移率变动分析进行检测。提取物中的蛋白质选择性地识别具有临床活性的铂修饰DNA片段。未观察到与反式顺铂修饰的DNA片段有结合。这些蛋白质可能在细胞毒性或DNA修复过程中发挥作用。
