Diallyl trisulfide suppresses the proliferation and induces apoptosis of human colon cancer cells through oxidative modification of beta-tubulin.
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 microM, DLD-1 IC50 = 13.3 microM); 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 G1 and S phases were decreased by DATS, and their populations at the G2/M phase were markedly increased for up to 12 h. The cells with a sub-G1 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 G1/S boundary revealed cell cycle-dependent induction of apoptosis through the transition of the G2/M phase to the G1 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-12beta and Cys-354beta 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.
Hosono
Takashi
T
Department of Applied Life Sciences, Nihon University Graduate School of Bioresource Sciences, Kanagawa 252-8510, Japan.
Fukao
Tomomi
T
Ogihara
Jun
J
Ito
Yoshimasa
Y
Shiba
Hajime
H
Seki
Taiichiro
T
Ariga
Toyohiko
T
D000498:Allyl Compounds / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D015153:Blotting, Western; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002469:Cell Separation; D002474:Cell-Free System; D002853:Chromatography, Liquid; D003110:Colonic Neoplasms / Q000378:metabolism*; D019926:Cyclin B / Q000378:metabolism; D056744:Cyclin B1; D003545:Cysteine / Q000031:analogs & derivatives / Q000737:chemistry; D003593:Cytoplasm / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D005737:Garlic; D006801:Humans; D020128:Inhibitory Concentration 50; D051379:Mice; D008819:Mice, Nude; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure; D009368:Neoplasm Transplantation; D018384:Oxidative Stress; D010100:Oxygen / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D013440:Sulfides / Q000737:chemistry / Q000494:pharmacology*; D013997:Time Factors; D014404:Tubulin / Q000737:chemistry*
D000498:Allyl Compounds; D000970:Antineoplastic Agents; C534699:CCNB1 protein, human; C534700:Ccnb1 protein, mouse; D019926:Cyclin B; D056744:Cyclin B1; D004220:Disulfides; D010455:Peptides; C086300:S-allylmercaptocysteine; D013440:Sulfides; D014404:Tubulin; C042577:diallyl trisulfide; C028009:diallyl disulfide; D004247:DNA; D003545:Cysteine; D010100:Oxygen