Diallyl disulfide (DADS) induced apoptosis undergo caspase-3 activity in human bladder cancer T24 cells.
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.
Lu
H F
HF
Department of Clinical Pathology, Cheng-Hsin Rehabilitation Medical Center, Taipei 100, Taiwan, ROC.
Sue
C C
CC
Yu
C S
CS
Chen
S C
SC
Chen
G W
GW
Chung
J G
JG
D000498:Allyl Compounds / Q000494:pharmacology*; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000911:Antibodies, Monoclonal; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D042846:CDC2-CDC28 Kinases / Q000378:metabolism; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D045744:Cell Line, Tumor; D002470:Cell Survival; D051357:Cyclin-Dependent Kinase 2; D053938:DNA Fragmentation; D004273:DNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D004220:Disulfides / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007202:Indicators and Reagents; D020411:Oligonucleotide Array Sequence Analysis; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014343:Trypan Blue; D001749:Urinary Bladder Neoplasms / Q000473:pathology*
D000498:Allyl Compounds; D000590:Amino Acid Chloromethyl Ketones; D000911:Antibodies, Monoclonal; D000972:Antineoplastic Agents, Phytogenic; D004273:DNA, Neoplasm; D004220:Disulfides; D007202:Indicators and Reagents; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D011064:Poly Adenosine Diphosphate Ribose; C028009:diallyl disulfide; D006861:Hydrogen Peroxide; D042846:CDC2-CDC28 Kinases; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D014343:Trypan Blue