School of Korean Medicine, Pusan National University, Yangsan 626-870, Republic of Korea.
Anti-Aging Research Center & Blue-Bio Industry Regional Innovation Center, Dongeui University, Busan 614-714, Republic of Korea.
Food Chem Toxicol. 2014 Feb;64:157-65. doi: 10.1016/j.fct.2013.11.034. Epub 2013 Dec 1.
The present study was undertaken to determine whether sulforaphane-derived reactive oxygen species (ROS) might cause growth arrest and apoptosis in human bladder cancer 5637 cells. Our results show that the reduced viability of 5637 cells by sulforaphane is due to mitotic arrest, but not the G2 phase. The sulforaphane-induced mitotic arrest correlated with an induction of cyclin B1 and phosphorylation of Cdk1, as well as a concomitant increased complex between cyclin B1 and Cdk1. Sulforaphane-induced apoptosis was associated with the activation of caspase-8 and -9, the initiators caspases of the extrinsic and intrinsic apoptotic pathways, respectively, and activation of effector caspase-3 and cleavage of poly (ADP-ribose) polymerase. However, blockage of caspase activation inhibited apoptosis and abrogated growth inhibition in sulforaphane-treated 5637 cells. This study further investigated the roles of ROS with respect to mitotic arrest and the apoptotic effect of sulforaphane, and the maximum level of ROS accumulation was observed 3h after sulforaphane treatment. However, a ROS scavenger, N-acetyl-L-cysteine, notably attenuated sulforaphane-mediated apoptosis as well as mitotic arrest. Overall, these results suggest that sulforaphane induces mitotic arrest and apoptosis of 5637 cells via a ROS-dependent pathway.
本研究旨在确定萝卜硫素衍生的活性氧（ROS）是否可能导致人膀胱癌 5637 细胞生长停滞和凋亡。我们的结果表明，萝卜硫素降低 5637 细胞活力是由于有丝分裂阻滞，而不是 G2 期。萝卜硫素诱导的有丝分裂阻滞与细胞周期蛋白 B1 的诱导和 Cdk1 的磷酸化相关，以及细胞周期蛋白 B1 和 Cdk1 之间的复合物同时增加。萝卜硫素诱导的细胞凋亡与 caspase-8 和 caspase-9 的激活相关，分别为外源性和内源性凋亡途径的起始半胱氨酸蛋白酶，以及效应半胱氨酸蛋白酶-3 的激活和多聚（ADP-核糖）聚合酶的切割。然而，半胱氨酸天冬氨酸蛋白酶激活抑制剂抑制 caspase 激活可抑制凋亡，并消除萝卜硫素处理的 5637 细胞中的生长抑制。本研究进一步探讨了 ROS 对有丝分裂阻滞和萝卜硫素凋亡作用的作用，并且在萝卜硫素处理后 3 小时观察到 ROS 积累的最大水平。然而，ROS 清除剂 N-乙酰-L-半胱氨酸显著减弱了萝卜硫素介导的凋亡以及有丝分裂阻滞。总的来说，这些结果表明萝卜硫素通过 ROS 依赖性途径诱导 5637 细胞的有丝分裂阻滞和凋亡。
