Lu H F, Sue C C, Yu C S, Chen S C, Chen G W, Chung J G
Department of Clinical Pathology, Cheng-Hsin Rehabilitation Medical Center, Taipei 100, Taiwan, ROC.
Food Chem Toxicol. 2004 Oct;42(10):1543-52. doi: 10.1016/j.fct.2003.06.001.
Diallyl disulfide (DADS), one of the major components of garlic (Allium sativum), is well known to have chemopreventative activity against human cancer such as colon, lung and skin. But the exact mechanism of the action is still unclear. In this study, we investigated how DADS--induced cell cycle arrest and apoptosis in T24 human bladder cancer cells in vitro. Apoptosis induction, cell viability, cell cycle arrest, caspases-3, -9 activity and gene expression were measured to determine their variation by flow cytometric assay, western blot, and determination of caspase-3 activity, PCR and cDNA microarray. There are significant differences in cell death (decreased viable cells then increased the amounts of apoptosis) of T24 cells that were detected between DADS (5-75 microM) treated and untreated groups. A significant increase was found in apoptosis induction when cells were treated with DADS (50 microM) compared to without DADS treated groups. DADS also promoted caspase-3 activity after exposure for 1, 3, 6, 12, and 24 h, which led to induce apoptosis. DADS also increased the product of intracellular hydrogen peroxide. Furthermore, the DADS-induced apoptosis on T24 cells was blocked by the broad-spectrum caspase inhibitor, z-VAD-fmk and antioxidant (catalase). DADS also increased cyclin E and decreased CDK2 gene expression which may lead to the G2/M arrest of T24 cells.
二烯丙基二硫化物(DADS)是大蒜(葱属植物)的主要成分之一,众所周知它对结肠癌、肺癌和皮肤癌等人类癌症具有化学预防活性。但其确切作用机制仍不清楚。在本研究中,我们在体外研究了DADS如何诱导T24人膀胱癌细胞的细胞周期停滞和凋亡。通过流式细胞术、蛋白质印迹法以及半胱天冬酶-3活性测定、聚合酶链反应和cDNA微阵列检测来测量凋亡诱导、细胞活力、细胞周期停滞、半胱天冬酶-3、-9活性及基因表达,以确定它们的变化。在DADS(5-75微摩尔)处理组和未处理组之间检测到的T24细胞的细胞死亡(活细胞减少,随后凋亡细胞数量增加)存在显著差异。与未用DADS处理的组相比,用DADS(50微摩尔)处理细胞时凋亡诱导显著增加。DADS在暴露1、3、6、12和24小时后还促进了半胱天冬酶-3的活性,这导致诱导凋亡。DADS还增加了细胞内过氧化氢的产生。此外,广谱半胱天冬酶抑制剂z-VAD-fmk和抗氧化剂(过氧化氢酶)可阻断DADS诱导的T24细胞凋亡。DADS还增加了细胞周期蛋白E的表达并降低了细胞周期蛋白依赖性激酶2(CDK2)的基因表达,这可能导致T24细胞的G2/M期停滞。