Chen Ping-Ping, Li Chao-Yi, Han Yu, Chen Xue-Yan, Liu Huan-Long, Du Yu-Min, Su Su-Wen, Zhang Yong-Jian
aThe Key Laboratory of Neural and Vascular Biology, Ministry of Education bThe Key Laboratory of Pharmacology and Toxicology for New Drugs, Department of Pharmacology cThe Department of Pharmaceutical Chemistry, Hebei Medical University dPharmaceutical Department of the Second Hospital of Hebei Medical University, Shijiazhuang, Hebei Province, China.
Anticancer Drugs. 2015 Jul;26(6):620-31. doi: 10.1097/CAD.0000000000000226.
N-[4-(4,6-Dimethyl-2-pyrimidinyloxy)-3-methylphenyl]-N'-[2-(dimethylamino)]benzoylurea (SUD) is a novel synthesized benzoylurea derivative. We selected several human cancer cell lines to investigate whether SUD can inhibit the growth of cancer cells. We selected the liver cell line L-02 to investigate the effect of SUD on the normal cells. Flow cytometric analysis was used to detect the effect of SUD on cell cycle, Hoechst 33258 staining was used to evaluate the apoptosis induced by SUD, real-time fluorescence quantitative PCR was used to investigate the expression of the cell cycle-relevant and apoptosis-relevant genes, a reactive oxygen species (ROS) assay was used to observe the production of ROS, and western blotting was used to determine the level of cell cycle-relevant and apoptosis-relevant proteins. According to the results of the MTT assay, the growth of human cancer cell lines was significantly inhibited by SUD treatment in a time-dependent and concentration-dependent manner; however, the growth of human normal cells was not significantly inhibited by SUD treatment. The results of flow cytometric analyses showed that SUD induced cell-cycle arrest at the G2-phase in MCF-7 cells and at the G1-phase in BGC-823 cells. The results of Hoechst 33258 staining showed that SUD induced apoptosis in MCF-7 and BGC-823 cells. The results of the ROS assay showed that the production of ROS was increased by SUD in MCF-7 and BGC-823 cells. Our research suggests that the growth-inhibitory effect of SUD on MCF-7 cells was related to G2-phase arrest, which was associated with the upregulated expression of p53 and Chk1 proteins, and downregulation of the cyclin B1 gene, cdc25a, and cyclin-dependent kinase 1 (CDK1) proteins; the growth-inhibitory effect of SUD on BGC-823 cells was related to G1-phase arrest, which was associated with upregulation of the p53 gene and Chk1 protein and downregulation of cdc25a protein and the CDK4 gene. SUD also induced apoptosis in MCF-7 and BGC-823 cell lines through the mitochondrial pathway in a p53-dependent manner.
N-[4-(4,6-二甲基-2-嘧啶氧基)-3-甲基苯基]-N'-[2-(二甲基氨基)]苯甲酰脲（SUD）是一种新合成的苯甲酰脲衍生物。我们选择了几种人类癌细胞系来研究SUD是否能抑制癌细胞的生长。我们选择肝细胞系L-02来研究SUD对正常细胞的影响。采用流式细胞术分析检测SUD对细胞周期的影响，用Hoechst 33258染色评估SUD诱导的细胞凋亡，用实时荧光定量PCR研究细胞周期相关基因和凋亡相关基因的表达，用活性氧（ROS）检测法观察ROS的产生，并用蛋白质印迹法测定细胞周期相关蛋白和凋亡相关蛋白的水平。根据MTT检测结果，SUD处理以时间和浓度依赖性方式显著抑制人类癌细胞系的生长；然而，SUD处理并未显著抑制人类正常细胞的生长。流式细胞术分析结果显示，SUD诱导MCF-7细胞在G2期发生细胞周期阻滞，诱导BGC-823细胞在G1期发生细胞周期阻滞。Hoechst 33258染色结果显示，SUD诱导MCF-7和BGC-823细胞凋亡。ROS检测结果显示，SUD使MCF-7和BGC-823细胞中的ROS产生增加。我们的研究表明，SUD对MCF-7细胞的生长抑制作用与G2期阻滞有关，这与p53和Chk1蛋白表达上调以及细胞周期蛋白B1基因、cdc25a和细胞周期蛋白依赖性激酶1（CDK1）蛋白表达下调有关；SUD对BGC-823细胞的生长抑制作用与G1期阻滞有关，这与p53基因和Chk1蛋白表达上调以及cdc25a蛋白和CDK4基因表达下调有关。SUD还通过线粒体途径以p53依赖的方式诱导MCF-7和BGC-823细胞系凋亡
