新型噻唑并三嗪类化合物R001抑制ERK1/2磷酸化的抗肿瘤作用  

The Antitumor Activity of the Novel Thiazolo-Triazin Compound R001 in Inhibiting ERK1/2 Phosphorylation

作　　者：吴凤娟[1] Diwa Koirala 刘斯婕[2] 金辄[2] 胡春[2] 李大伟[1] 

机构地区：[1]上海交通大学药学院,上海200240 [2]沈阳药科大学制药工程学院,辽宁沈阳110016

出　　处：《现代生物医学进展》2012年第29期5638-5642,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金(21072130)

摘　　要：目的:探讨噻唑并三嗪类化合物R001对ERK1/2磷酸化的影响及其抗肿瘤活性。方法:采用基于U2OS-EGFP细胞的荧光分析法和MTT法检测化合物R001对U2OS-EGFP-4F12G和U2OS、T47D、SKOV3、MCF7等其他肿瘤细胞增殖的影响;采用电子显微镜观察化合物R001对MCF7、T47D、U2OS-EGFP-4F12G和SKOV3等多种肿瘤细胞株增殖的抑制作用;采用流式细胞术检测化合物R001对MCF7、U2OS-EGFP-4F12G和SKOV3等多种肿瘤细胞周期的影响;采用免疫印迹法检测检测化合物R001对ERK1/2磷酸化的影响。结果:基于U2OS-EGFP细胞的荧光分析法显示,化合物R001抑制U2OS-EGFP-4F12G细胞增殖的IC50值为11.58μM;MTT法显示,化合物R001对U2OS、T47D、SKOV3、MCF7等多种肿瘤细胞增殖的影响具有剂量依赖性;电子显微镜观察显示,化合物R001可以引起MCF7、SKOV3和U2OS-EGFP-4F12G三种肿瘤细胞的数量和形态变化;流式细胞术显示,化合物R001可将MCF7、U2OS-EGFP-4F12G和SKOV3等肿瘤细胞阻断在S期;免疫印迹试验显示,化合物R001可以有效抑制SKOV3和U2OS细胞中ERK1/2的磷酸化。结论:噻唑并三嗪类化合物R001通过抑制ERK1/2磷酸化而发挥抗肿瘤作用。ABSTRACT Objective： To investigate the effect of the novel kinase inhibitor thiazolo-triazin. Compound： R001 on ERK1/2 phosphorylation and its antitumor activity. Method： Fluorescence assay based on U2OS-EGFP-4F12G cell culture and MTT assay was used to measure the cell proliferation effect of Compound R001 on U2OS-EGFP-4F12G and U2OS, T47D, SKOV3 and MCF7 cell lines. Electron microscopic technique was used to observe the number and morphologic changes of MCF7, T47D, U2OS-EGFP-4F12G and SKOV3 cell lines. Flow cytometry was used to estimate the effect of the Compound R001 on the cell cycle in MCF7, U2OS-EGFP-4F12G and SKOV3 ceils with propidium iodide （PI）. Western blotting was used to detect the effect of Compound R001 on ERK1/2 phosphorylation in SKOV3 and U2OS cells. Results： Fluorescence assay based on U2OS-EGFP-4F12G cell culture indicated the median inhibitory concentration （IC50） value of Compound R001 was 11.58 IxM. MTT assay demonstrated that Compound R001 could inhibit U2OS, T47D, SKOV3 and MCF7 cells proliferation dose-dependently. The results of electron microscopic showed Compound R001 reduced the cell number and induced cellular morphology changes of MCF7, SKOV3 and U2OS-EGFP-4F12G cells. FACS （Analysis of cell cycle） revealed that MCF7, U2OS-EGFP-4F12G and SKOV3 were induced to arrest at phase S while treated with Compound R001. Western blotting displayed that Compound R001 could inhibit the phosphorylation of ERK1/2 in both SKOV3 and U2OS cell lines distinctly. Conclusions： The novel thiazolo-triazin Compound R001 has an antitumor activity through inhibiting ERK1/2 phosphorylation. Key Words： Thiazolo-triazin compounds; ERK1/2 phosphorylation; Antitumor

关 键 词：噻唑并三嗪类化合物 ERK1 2磷酸化 抗肿瘤 

分 类 号：R915[医药卫生—微生物与生化药学] R730.5[医药卫生—药学]