Song Ju-Dong, Lee Sang Kwon, Kim Kang Mi, Park Si Eun, Park Sung-Joo, Kim Koan Hoi, Ahn Soon Cheol, Park Young Chul
Department of Microbiology and Immunology, Pusan National University School of Medicine, Busan, Republic of Korea.
J Biochem Mol Toxicol. 2009 Jan-Feb;23(1):71-9. doi: 10.1002/jbt.20266.
Diallyl disulfide (DADS) is the most prevalent oil-soluble sulfur compound in garlic and inhibits cell proliferation in many cancer cell lines. Here we examined DADS cytotoxicity in a redox-mediated process, involving reactive oxygen species (ROS) production. In the present study, p53-independent cell cycle arrest at G2/M phase was observed with DADS treatment, along with time-dependent increase of cyclin B1. In addition, apoptosis was also observed upon 24-h DADS treatment accompanied by activation of p53. In HCT-116 cells, DADS application induced a dose-dependent increase and time-dependent changes in ROS production. Scavenging of DADS-induced ROS by N-acetyl cysteine or reduced glutathione inhibited cell cycle arrest, apoptosis and p53 activation by DADS. These results suggest that ROS trigger the DADS-induced cell cycle arrest and apoptosis and that ROS are involved in stress-induced signaling upstream of p53 activation. Transfection of p53 small interfering RNA prevents the accumulation of cleaved poly(ADP-ribose) polymerase and sub-G1 cell population by 65% and 35%, respectively. Moreover, DADS-induced apoptosis was also prevented by treatment with oligomycin, which is known to prevent p53-dependent apoptosis by reducing ROS levels in mitochondria. These results suggest that mitochondrial ROS may serve as second messengers in DADS-induced apoptosis, which requires activation of p53.
二烯丙基二硫化物（DADS）是大蒜中最普遍的油溶性硫化合物，可抑制多种癌细胞系中的细胞增殖。在此，我们研究了DADS在涉及活性氧（ROS）产生的氧化还原介导过程中的细胞毒性。在本研究中，用DADS处理后观察到p53非依赖性细胞周期在G2/M期停滞，同时细胞周期蛋白B1随时间增加。此外，在24小时DADS处理后还观察到凋亡，同时伴有p53激活。在HCT-116细胞中，应用DADS诱导ROS产生呈剂量依赖性增加和时间依赖性变化。用N-乙酰半胱氨酸或还原型谷胱甘肽清除DADS诱导的ROS可抑制DADS诱导的细胞周期停滞、凋亡和p53激活。这些结果表明，ROS触发DADS诱导的细胞周期停滞和凋亡，并且ROS参与p53激活上游的应激诱导信号传导。转染p53小干扰RNA可分别使裂解的聚（ADP-核糖）聚合酶和亚G1细胞群体的积累减少65%和35%。此外，用寡霉素处理也可防止DADS诱导的凋亡，已知寡霉素可通过降低线粒体中的ROS水平来防止p53依赖性凋亡。这些结果表明，线粒体ROS可能作为DADS诱导凋亡的第二信使，这需要p53激活。
