Institute of Clinical Medical Science, China Medical University, Taichung, Taiwan; School of Dentistry, China Medical University, Taichung, Taiwan.
School of Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung, Taiwan; China Medical University Children's Hospital, China Medical University, Taichung, Taiwan.
J Formos Med Assoc. 2021 Jan;120(1 Pt 2):515-523. doi: 10.1016/j.jfma.2020.06.025. Epub 2020 Jul 3.
BACKGROUND/PURPOSE: Cisplatin-resistant oral cancer is clinically difficult to manage and the dose-dependent toxicities of cisplatin has been widely concerned. Allyl isothiocyanate (AITC), known as mustard oil, is a plant-derived compound abundant in cruciferous vegetables. It is reported to have anti-cancer potential as a natural dietary chemopreventive compound against a variety of cancers, but the effect of AITC on cisplatin-resistant cancer cells is still little-known.
Human CAL27-cisplatin-resistant oral cancer cells (CAR cells) were examined to investigate the antitumor properties of AITC. 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay, IncuCyte™ S3 cell proliferation assay, 4',6-diamidino-2-phenylindole (DAPI) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining as well as Western blot analysis were deployed.
AITC decreased CAR cell viability, induced cell death of CAR cells and inhibited the confluences of cultured CAR cells. When CAR cells were treated with AITC, activation of caspase-3 and caspase-9 by AITC was observed and could be reversed by Z-VAD-fmk (pan-caspase inhibitor). Furthermore, the protein expressions of phosphorylated protein kinase B (p-AKT) and phosphorylated mammalian target of rapamycin (p-mTOR) were suppressed in AITC-treated CAR cells, whereas protein expressions of Bax, cytochrome c, Apaf-1, cleaved caspase-3, and cleaved caspase-9 were upregulated in AITC-treated CAR cells.
AITC can inhibit Akt/mTOR proliferation signaling and promote mitochondria-dependent apoptotic pathway through AITC-enhanced activities of caspase-3 and caspase-9 in CAR cells.
背景/目的：顺铂耐药口腔癌临床治疗困难，顺铂的剂量依赖性毒性已引起广泛关注。丙烯基异硫氰酸酯（AITC），又称芥末油，是一种在十字花科蔬菜中含量丰富的植物衍生化合物。据报道，作为一种天然的膳食化学预防化合物，AITC 具有抗癌潜力，可预防多种癌症，但 AITC 对顺铂耐药癌细胞的作用仍知之甚少。
检测人 CAL27-顺铂耐药口腔癌细胞（CAR 细胞），以研究 AITC 的抗肿瘤特性。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测法、IncuCyte™ S3 细胞增殖检测法、4',6-二脒基-2-苯基吲哚（DAPI）和末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）染色以及 Western blot 分析等方法。
AITC 降低了 CAR 细胞活力，诱导 CAR 细胞死亡，并抑制培养的 CAR 细胞的融合。当 CAR 细胞用 AITC 处理时，观察到 AITC 激活了 caspase-3 和 caspase-9，而 Z-VAD-fmk（泛 caspase 抑制剂）可逆转这种激活。此外，AITC 处理的 CAR 细胞中磷酸化蛋白激酶 B（p-AKT）和磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）的蛋白表达受到抑制，而 AITC 处理的 CAR 细胞中 Bax、细胞色素 c、Apaf-1、cleaved caspase-3 和 cleaved caspase-9 的蛋白表达上调。
AITC 可通过增强 CAR 细胞中 caspase-3 和 caspase-9 的活性，抑制 Akt/mTOR 增殖信号通路，并促进线粒体依赖性凋亡途径。
