UPM-MAKNA Cancer Research Laboratory, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
CREA Consiglio per la ricerca in agricoltura e l'analisi dell'economia agrarian, Centro di ricerca Agricoltura e Ambiente (CREA-AA), Via di Corticella 133, 40128 Bologna, Italy.
Molecules. 2018 Nov 27;23(12):3092. doi: 10.3390/molecules23123092.
Glucoraphasatin (GRH), a glucosinolate present abundantly in the plants of the family, is hydrolyzed by myrosinase to raphasatin, which is considered responsible for its cancer chemopreventive activity; however, the underlying mechanisms of action have not been investigated, particularly in human cell lines. The aims of this study are to determine the cytotoxicity of raphasatin, and to evaluate its potential to cause apoptosis and modulate cell cycle arrest in human breast adenocarcinoma MCF-7 cells. The cytotoxicity was determined following incubation of the cells with glucoraphasatin or raphasatin (0⁻100 µM), for 24, 48, and 72 h. GRH displayed no cytotoxicity as exemplified by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. When myrosinase was added to the incubation system to convert GRH to raphasatin, cytotoxicity was evident. Exposure of the cells to raphasatin stimulated apoptosis, as was exemplified by cell shrinkage, membrane blebbing, chromatin condensation, and nuclear fragmentation. Moreover, using Annexin V-FITC assay, raphasatin induced apoptosis, as witnessed by changes in cellular distribution of cells, at different stages of apoptosis; in addition, raphasatin caused the arrest of the MCF-7 cells at the G₂ + M phase. In conclusion, raphasatin demonstrated cancer chemopreventive potential against human breast adenocarcinoma (MCF-7) cells, through induction of apoptosis and cell cycle arrest.
葡萄糖硫苷（GRH）是十字花科植物中大量存在的一种硫代葡萄糖苷，被黑芥子酶水解为莱菔硫烷，被认为是其抗癌化学预防活性的原因；然而，其作用机制尚未得到研究，特别是在人细胞系中。本研究的目的是确定莱菔硫烷的细胞毒性，并评估其在人乳腺癌 MCF-7 细胞中诱导细胞凋亡和调节细胞周期停滞的潜力。用葡萄糖硫苷或莱菔硫烷（0⁻100 µM）孵育细胞 24、48 和 72 h 后，测定细胞毒性。3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法表明，GRH 没有细胞毒性。当黑芥子酶被添加到孵育系统中将 GRH 转化为莱菔硫烷时，细胞毒性是明显的。细胞暴露于莱菔硫烷刺激细胞凋亡，如细胞皱缩、膜起泡、染色质浓缩和核碎裂。此外，用 Annexin V-FITC 测定法，莱菔硫烷诱导细胞凋亡，如细胞在凋亡不同阶段的细胞分布变化所示；此外，莱菔硫烷导致 MCF-7 细胞停滞在 G₂ + M 期。总之，莱菔硫烷通过诱导细胞凋亡和细胞周期停滞，显示出对人乳腺癌（MCF-7）细胞的抗癌化学预防潜力。
