Diallyl trisulfide suppresses the proliferation and induces apoptosis of human colon cancer cells through oxidative modification of β-tubulin
Diallyl trisulfide suppresses the proliferation and induces apoptosis of human colon cancer cells through oxidative modification of β-tubulin
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DOI：
10.1074/jbc.m507127200
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发表时间：
2005-12-16
影响因子：
4.8
通讯作者：
Ariga, T
中科院分区：
文献类型：
--
作者：
Hosono, T;Fukao, T;Ariga, T
烯丙基硫醚是从大蒜中获得的特征风味成分。这些硫化物被认为是负责其流行病学证明抗癌作用的大蒜食用者。本研究以人结肠癌细胞系HCT-15和DLD-1为研究对象，探讨大蒜抗肿瘤作用的分子基础。二烯丙基三硫化物显著抑制细胞的生长（DATS，HCT-15 IC 50 = 11.5 μ M，DLD-1 IC 50 = 13.3 μ M）;然而，二烯丙基一硫化物和二烯丙基二硫化物均未显示这种作用。DATS可明显降低HCT-15和DLD-1细胞在G1期和S期的比例，增加G2/M期细胞的比例，作用时间可达12 h。具有sub-G（1）DNA含量的细胞此后增加。Caspase-3活性也显著增加DATS。对停滞在G（1）/S边界的细胞进行的细胞凋亡激活细胞分选仪分析显示，通过DATS将G（2）/M期转变为G（1）期，细胞周期依赖性诱导凋亡。在体外无细胞系统中，DATS抑制微管蛋白聚合。DATS破坏了细胞微管网络的形成，在间期可以看到微管碎片。通过液相色谱-串联质谱法对DATS处理的微管蛋白进行肽质量图谱分析表明，半胱氨酸残基Cys-12 β和Cys-354 β发生特异性氧化修饰，形成S-烯丙基巯基半胱氨酸，肽质量增加72.1 Da。DATS在携带HCT-15异种移植物的裸鼠中也表现出了有效的抗肿瘤活性。这是第一篇描述大蒜成分直接修饰细胞内靶分子的论文。
Allyl sulfides are characteristic flavor components obtained from garlic. These sulfides are thought to be responsible for their epidemiologically proven anticancer effect on garlic eaters. This study was aimed at clarifying the molecular basis of this anticancer effect of garlic by using human colon cancer cell lines HCT-15 and DLD-1. The growth of the cells was significantly suppressed by diallyl trisulfide (DATS, HCT-15 IC50 = 11.5 mu M, DLD-1 IC50 = 13.3 mu M); however, neither diallyl monosulfide nor diallyl disulfide showed such an effect. The proportion of HCT-15 and that of DLD-1 cells residing at the G(1) and S phases were decreased by DATS, and their populations at the G(2)/M phase were markedly increased for up to 12 h. The cells with a sub-G(1) DNA content were increased thereafter. Caspase-3 activity was also dramatically increased by DATS. Fluorescence-activated cell sorter analysis performed on the cells arrested at the G(1)/S boundary revealed cell cycle-dependent induction of apoptosis through the transition of the G(2)/M phase to the G(1) phase by DATS. DATS inhibited tubulin polymerization in an in vitro cell-free system. DATS disrupted microtubule network formation of the cells, and microtubule fragments could be seen at the interphase. Peptide mass mapping by liquid chromatography-tandem mass spectrometry analysis for DATS-treated tubulin demonstrated that there was a specific oxidative modification of cysteine residues Cys-12 beta and Cys-354 beta to form S-allylmercaptocysteine with a peptide mass increase of 72.1 Da. The potent antitumor activity of DATS was also demonstrated in nude mice bearing HCT-15 xenografts. This is the first paper describing intracellular target molecules directly modified by garlic components.