Gomez-Verjan Juan C, Estrella-Parra Edgar A, González-Sánchez Ignacio, Vázquez-Martínez Edgar Ricardo, Vergara-Castañeda Edgar, Cerbón Marco A, Reyes-Chilpa Ricardo
Departamento de Productos Naturales, Instituto de Química, Universidad Nacional Autónoma de México, México City, Distrito Federal, México; Departamento de Biología, Facultad de Química, Universidad Nacional Autónoma de México, México City, Distrito Federal, México.
J Pharm Pharmacol. 2014 Aug;66(8):1189-95. doi: 10.1111/jphp.12245. Epub 2014 Mar 27.
The aim of this study was to determine the cellular and molecular mechanisms of cell death induced by mammea A/BA and A/BB (3 : 1) on K562 cells.
These compounds were isolated from Calophyllum brasiliense and its cytotoxicity was tested using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell death was evaluated by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and immunocytofluorescence of active caspase-3. Genotoxicity was tested using comet assay. Lastly, a chemoinformatic analysis was performed with Osiris-Molinspiration software.
The mixture of mammea A/BA and A/BB (3 : 1) showed cytotoxic activity against K562 cells (IC50 = 43.5 μm). TUNEL positive cells and active caspase-3 were detected after treatment. Genotoxicity of mammea A/BA and A/BB on K562 was detected since first hour of treatment. Additionally, mammea A/BA and A/BB were found to be in compliance with Lipinski 'rule of 5' suggesting that they possess strong potential of druglikeness.
The overall results confirm and extend the knowledge about coumarins as an important resource of antitumor drugs, and indicate that these compounds could be used in further preclinical studies against leukaemia.
本研究旨在确定巴西红厚壳中的 mamm ea A/BA 和 A/BB（3∶1）诱导 K562 细胞死亡的细胞和分子机制。
从巴西红厚壳中分离出这些化合物，并使用 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐法检测其细胞毒性。通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）法和活性半胱天冬酶-3 的免疫细胞荧光法评估细胞死亡情况。使用彗星试验检测遗传毒性。最后，使用 Osiris-Molinspiration 软件进行化学信息学分析。
mammea A/BA 和 A/BB（3∶1）的混合物对 K562 细胞具有细胞毒性活性（IC50 = 43.5 μ m）。处理后检测到 TUNEL 阳性细胞和活性半胱天冬酶-3。从处理的第一小时起就检测到了 mamm ea A/BA 和 A/BB 对 K562 的遗传毒性。此外，发现 mamm ea A/BA 和 A/BB 符合 Lipinski 的“五规则”，表明它们具有很强的类药物潜力。
总体结果证实并扩展了关于香豆素作为抗肿瘤药物重要来源的知识，并表明这些化合物可用于进一步针对白血病的临床前研究。
