Department of Biochemistry, Dongeui University College of Korean Medicine, Busan 47227, Republic of Korea; Anti-Aging Research Center & Blue-Bio Industry RIC, Dongeui University, Busan 47340, Republic of Korea.
Biomed Pharmacother. 2017 Oct;94:63-71. doi: 10.1016/j.biopha.2017.07.055. Epub 2017 Jul 26.
Diallyl trisulfide (DATS), one of the principal constituents of garlic oil, is a kind of organosulfur compound with high anti-cancer activity. Although inhibition of cancer cell proliferation by DATS is known to be associated with the induction of apoptosis and cell cycle arrest related to reactive oxygen species (ROS) production, it is still necessary to study the detailed mechanisms. In this study, we investigated the role of ROS on the activation of AMP-activated protein kinase (AMPK) in DATS-induced apoptosis and cell cycle arrest in AGS human gastric carcinoma cells. The results of the present study indicate that DATS inhibited proliferation of AGS cells by promoting apoptosis, and accumulating cellular portion of G2/M phase via the induction of cyclin B1 and cyclin-dependent kinase p21(WAF1/CIP1). The phosphorylation of histone H3 was also markedly increased following treatment with DATS, revealing that DATS stimulated a mitotic arrest, not the G2 phase. Furthermore, we found that DATS concurrently induced phosphorylation of AMPK; however, chemical inhibition of AMPK by compound C, an AMPK inhibitor, significantly blocked apoptosis induced by DATS, suggesting that DATS induces cytotoxicity of AGS cells through the AMPK-dependent pathway. Moreover, DATS provoked intracellular ROS generation and the loss of mitochondrial membrane potential, and in particular, when ROS production was blocked by antioxidant N-acety-l-cysteine, both AMPK activation and growth inhibition by DATS were completely abolished. Collectively, these findings suggest that DATS inhibited growth of AGS cells, which was mediated by complex interplay between cellular mechanisms governing redox homeostasis, apoptosis, and cell cycle arrest, through a ROS-dependent activation of AMPK pathway.
二烯丙基三硫醚（DATS）是大蒜油的主要成分之一，是一种具有高抗癌活性的有机硫化合物。尽管 DATS 抑制癌细胞增殖与诱导与活性氧（ROS）产生相关的细胞凋亡和细胞周期停滞有关，但仍需要研究其详细的机制。在这项研究中，我们研究了 ROS 在 DATS 诱导的 AGS 人胃癌细胞凋亡和细胞周期停滞中激活 AMP 激活蛋白激酶（AMPK）中的作用。本研究结果表明，DATS 通过促进细胞凋亡和诱导细胞周期停滞于 G2/M 期，从而抑制 AGS 细胞的增殖，同时还诱导 cyclin B1 和细胞周期依赖性激酶 p21（WAF1/CIP1）的积累。DATS 处理后组蛋白 H3 的磷酸化也明显增加，表明 DATS 刺激有丝分裂停滞，而不是 G2 期。此外，我们发现 DATS 同时诱导 AMPK 的磷酸化；然而，用 AMPK 抑制剂化合物 C 化学抑制 AMPK 可显著阻断 DATS 诱导的细胞凋亡，表明 DATS 通过 AMPK 依赖途径诱导 AGS 细胞的细胞毒性。此外，DATS 引发细胞内 ROS 生成和线粒体膜电位丧失，特别是当抗氧化剂 N-乙酰半胱氨酸阻断 ROS 生成时，DATS 引起的 AMPK 激活和生长抑制完全被消除。综上所述，这些发现表明 DATS 通过 ROS 依赖的 AMPK 通路抑制 AGS 细胞的生长，这是通过调节氧化还原平衡、细胞凋亡和细胞周期停滞的细胞机制之间的复杂相互作用介导的。
