Lynch James T, Cockerill Mark J, Hitchin James R, Wiseman Daniel H, Somervaille Tim C P
Cancer Research UK Leukaemia Biology Laboratory, Paterson Institute for Cancer Research, University of Manchester, Manchester M20 4BX, UK.
Anal Biochem. 2013 Nov 1;442(1):104-6. doi: 10.1016/j.ab.2013.07.032. Epub 2013 Jul 31.
There is a lack of rapid cell-based assays that read out enzymatic inhibition of the histone demethylase LSD1 (lysine-specific demethylase 1). Through transcriptome analysis of human acute myeloid leukemia THP1 cells treated with a tranylcypromine-derivative inhibitor of LSD1 active in the low nanomolar range, we identified the cell surface marker CD86 as a sensitive surrogate biomarker of LSD1 inhibition. Within 24h of enzyme inhibition, there was substantial and dose-dependent up-regulation of CD86 expression, as detected by quantitative polymerase chain reaction, flow cytometry, and enzyme-linked immunosorbent assay. Thus, the use of CD86 expression may facilitate screening of compounds with putative LSD1 inhibitory activities in cellular assays.
目前缺乏能够读出对组蛋白去甲基化酶LSD1（赖氨酸特异性去甲基化酶1）酶抑制作用的基于细胞的快速检测方法。通过对用低纳摩尔范围内具有活性的反苯环丙胺衍生物LSD1抑制剂处理的人急性髓系白血病THP1细胞进行转录组分析，我们确定细胞表面标志物CD86是LSD1抑制作用的敏感替代生物标志物。在酶抑制24小时内，通过定量聚合酶链反应、流式细胞术和酶联免疫吸附测定法检测到CD86表达有显著的剂量依赖性上调。因此，利用CD86表达可能有助于在细胞检测中筛选具有假定LSD1抑制活性的化合物。
