Center for Molecular Systems Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Acta Pharmacol Sin. 2013 Jun;34(6):805-10. doi: 10.1038/aps.2013.61.
To identify novel small compound inhibitor of p53 protein.
Mouse embryonic fibroblasts (MEF) and mouse embryonic stem (ES) cells were tested. Cell proliferation rate was determined using a Cell Proliferation Kit. The mRNA and protein levels of p53-related genes were measured using real-time PCR and Western blotting, respectively. Global response in the p53 signaling network was analyzed using Illumina whole-genome expression BeadChips.
Treatment of MEF cells with a small molecule 1,4-bis-[4-(3-phenoxy-propoxy)-but-2-ynyl]-piperazine (G5) at 10 μmol/L for 24 h markedly reduced the mRNA and protein levels of the p53 downstream genes MDM2 and p21. In G5-treated ES cells, a total of 372 differentially expressed genes were identified, and 18 among them were direct downstream genes of p53; 6 out of 9 p53-repressed genes were upregulated, and 5 out of 9 p53-activated genes were downregulated. In both MEF cells and ES cells, treatment of with G5 (10 μmol/L) up to 48 h neither affected the proliferation rate nor caused morphological alterations.
G5 inhibits p53 activity and simultaneously preserves the normal growth and proliferation of cells, therefore is a new compound for studies of p53-mediated cell manipulation.
鉴定新型 p53 蛋白小分子抑制剂。
检测小鼠胚胎成纤维细胞（MEF）和小鼠胚胎干细胞（ES）细胞。使用细胞增殖试剂盒测定细胞增殖率。分别采用实时 PCR 和 Western blot 法测定 p53 相关基因的 mRNA 和蛋白水平。采用 Illumina 全基因组表达 BeadChips 分析 p53 信号网络的整体反应。
用小分子 1,4-双-[4-(3-苯氧基-丙氧基)-2-丁炔基]-哌嗪（G5）处理 MEF 细胞，浓度为 10 μmol/L，处理 24 h 后，明显降低了 p53 下游基因 MDM2 和 p21 的 mRNA 和蛋白水平。在 G5 处理的 ES 细胞中，共鉴定出 372 个差异表达基因，其中 18 个是 p53 的直接下游基因；9 个 p53 抑制基因中有 6 个上调，9 个 p53 激活基因中有 5 个下调。在 MEF 细胞和 ES 细胞中，用 G5（10 μmol/L）处理 48 h 既不影响细胞增殖率，也不引起形态改变。
G5 抑制 p53 活性，同时保持细胞的正常生长和增殖，因此是一种用于研究 p53 介导的细胞操作的新型化合物。
