Department of Biological Science, Gachon University of Medicine and Science, 534-2 Yeonsu-dong, Yeonsu-gu, Incheon 406-799, Republic of Korea.
Food Chem Toxicol. 2010 Aug-Sep;48(8-9):2259-64. doi: 10.1016/j.fct.2010.05.058. Epub 2010 May 25.
This study aimed to investigate the effects of fisetin, a common dietary natural flavonoid, on apoptosis of Huh-7 cells. The MTT assay was used to evaluate the reduction of cell viability. In the DNA fragmentation assay and comet assay, fisetin-induced DNA damage was visible as a formation of DNA fragmentation and comet tails. Fisetin also induced intracellular accumulation of reactive oxygen species. Two-dimensional gel electrophoresis was performed to evaluate protein expression in fisetin-treated and control cells, and Baculoviral IAP repeat-containing protein 8 (BIRC8) and apoptosis regulator Bcl-W (Bcl2L2) were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Their expression was confirmed by western blotting. The anticancer effect of fisetin in Huh-7 cells may result from the down regulation of BIRC8 and the Bcl2L2.
本研究旨在探讨一种常见的膳食天然类黄酮——漆黄素对 Huh-7 细胞凋亡的影响。采用 MTT 法评估细胞活力降低情况。在 DNA 片段化检测和彗星实验中,可见到漆黄素诱导的 DNA 损伤表现为 DNA 片段化和彗星尾巴的形成。漆黄素还诱导细胞内活性氧的积累。通过二维凝胶电泳评估漆黄素处理和对照细胞中的蛋白质表达,并用基质辅助激光解吸/电离飞行时间质谱法鉴定出杆状病毒 IAP 重复蛋白 8(BIRC8)和凋亡调节因子 Bcl-W(Bcl2L2)。用 Western blot 进行表达验证。漆黄素在 Huh-7 细胞中的抗癌作用可能是通过下调 BIRC8 和 Bcl2L2 实现的。