Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, 250 Wu-Hsing Street, Taipei 110, Taiwan.
Naunyn Schmiedebergs Arch Pharmacol. 2010 Jul;382(1):43-50. doi: 10.1007/s00210-010-0519-4. Epub 2010 May 7.
The aim of this study was to investigate the potential applications of 5,5-diphenyl-2-thiohydantoin-N10 (DPTH-N10) in the treatment of human colon cancer. Subcultured human colon cancer cell line, COLO-205, was used for examining the antiproliferation effect of DPTH-N10 on colon cancer. Thymidine incorporation and cell count were conducted to examine the antiproliferation effect of DPTH-N10. Western blot analysis was performed to examine the protein levels of cell cycle-related proteins. DNA fragmentation assay was performed to examine the occurrence of apoptosis. DPTH-N10 at a range of concentrations (0-30 microM) inhibits the proliferation but did not cause the cell death of COLO-205, indicating that it may have an inhibitory effect on the cell proliferation in COLO-205. The apoptosis was observed in COLO-205 when the DPTH-N10 concentrations were higher than 30 muM. Western blot analysis showed that the protein level of the cell cycle inhibitory protein, p21, in COLO-205 increased after DPTH-N10 treatment. Immunoprecipitation showed that the formation of the cyclin-dependent kinase (CDK)2-p21 complex was increased in the DPTH-N10-treated COLO-205. Kinase assay further demonstrated that the CDK2 activity was decreased in the DPTH-N10-treated COLO-205. DPTH-N10 caused growth inhibition in COLO-205 by inhibiting DNA synthesis and activating apoptosis. The findings from our previous in vitro studies in DPTH-N10-induced anti-angiogenic effect and from the present in vitro studies in DPTH-N10-induced antiproliferation effect on colon cancer cell line strongly suggest the potential applications of DPTH-N10 in the treatment of human colon cancer.
本研究旨在探讨 5,5-二苯-2-硫代乙内酰脲-N10（DPTH-N10）在治疗人类结肠癌中的潜在应用。使用亚培养的人结肠癌细胞系 COLO-205 来检测 DPTH-N10 对结肠癌的增殖抑制作用。通过胸苷掺入和细胞计数来检测 DPTH-N10 的增殖抑制作用。通过 Western blot 分析来检测细胞周期相关蛋白的蛋白水平。通过 DNA 片段化分析来检测细胞凋亡的发生。DPTH-N10 在 0-30μM 的浓度范围内抑制 COLO-205 的增殖，但不会导致细胞死亡，这表明它可能对 COLO-205 的细胞增殖具有抑制作用。当 DPTH-N10 浓度高于 30μM 时，在 COLO-205 中观察到凋亡。Western blot 分析显示，DPTH-N10 处理后 COLO-205 中的细胞周期抑制蛋白 p21 的蛋白水平增加。免疫沉淀显示，DPTH-N10 处理后的 COLO-205 中 cyclin-dependent kinase（CDK）2-p21 复合物的形成增加。激酶测定进一步表明，DPTH-N10 处理后的 COLO-205 中 CDK2 活性降低。DPTH-N10 通过抑制 DNA 合成和激活凋亡来抑制 COLO-205 的生长。我们之前在 DPTH-N10 诱导的抗血管生成作用的体外研究以及本研究中 DPTH-N10 对结肠癌细胞系的增殖抑制作用的体外研究结果表明，DPTH-N10 具有治疗人类结肠癌的潜在应用。
