Abstract
Chemoprevention is regarded as one of the most promising and realistic approaches in the prevention of human cancer. Diallyl sulfide (DAS), an organosulfur component of garlic has been known for its chemopreventive activities against various cancers and also in recent years, numerous investigations have shown that sulfur-containing compounds induce apoptosis in multiple cell lines and experimental animals. Thus the present study was focused to elucidate the anticancerous effect and the mode of action of DAS against Colo 320 DM colon cancer cells. DAS induced apoptosis in Colo 320 DM cells was revealed by flow cytometer analysis and phosphatidyl serine exposure. DAS also promoted cell cycle arrest substantially at G2/M phase in Colo 320 DM cells. The production of reactive oxygen intermediates, which were examined by 2,7-dichlorodihydrofluorescein diacetate (H2DCF-DA), increased with time, after treatment with DAS. The activities of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) were decreased upon DAS treatment, which shows the antiproliferative and the cytotoxic effects, respectively. The expression of NF-κB was upregulated in DAS treated cells, compared to normal cells. Further, DAS promoted the expression of caspase-3 and suppression of Extracellular Regulatory Kinase-2 (ERK-2) activity in Colo 320 DM cells that was determined by Western blot analysis. In conclusion, DAS increased the production of ROS, caused cell cycle arrest, decreased cell proliferation and induced apoptosis in Colo 320 DM cells. Thus, this study put forward DAS as a drug that can possibly be used to treat cancers.
Similar content being viewed by others
References
Jass RJ (2002) Lower gastrointestinal tract. In: Alison M (ed) The cancer handbook, chap 38. Nature Publication Group, pp 543–561
Huerta S, Goulet EJ, Livingston EH (2006) Colon cancer and apoptosis. Am J Surg 191:517–526
Lakshman M, Subramaniam V, Jothy S (2004) CD44 negatively regulates apoptosis in murine colonic epithelium via the mitochondrial pathway. Exp Mol Pathol 76:196–204
Wyllie AH, Kerr JF, Currie AR (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68:251–306
Reed JC (2000) Mechanisms of apoptosis. Am J Pathol 157:1415–1430
Thornberry NA, Lazebnik Y (1998) Caspases: enemies within. Science 281:1312–1316
Thompson CB (1995) Apoptosis in the pathogenesis and treatment of disease. Science 267:1456–1462
Finkel T, Holbrook NJ (2000) Oxidants, oxidative stress and the biology of ageing. Nature 408:239–247
Szatrowski TP, Nathan CF (1991) Production of large amounts of hydrogen peroxide by human tumor cells. Cancer Res 51:794–798
Toyokuni S, Okamoto K, Yodoi J, Hiai H (1995) Persistent oxidative stress in cancer. FEBS Lett 358:1–3
Yan LJ, Levine RL, Sohal RS (1997) Oxidative damage during aging targets mitochondrial aconitase. Proc Natl Acad Sci USA 94:11168–11172
La Vecchia C, Tavani A (1998) Fruit and vegetables, and human cancer. Eur J Cancer Prev 7:3–8
Sengupta A, Ghosh S, Das S (2004) Modulatory influence of garlic and tomato on cyclooxygenase-2 activity, cell proliferation and apoptosis during azoxymethane induced colon carcinogenesis in rat. Cancer Lett 208:127–136
Ip C, Lisk DJ, Stoewsand GS (1992) Mammary cancer prevention by regular garlic and selenium-enriched garlic. Nutr Cancer 17:279–286
Sumiyoshi H, Wargovich MJ (1990) Chemoprevention of 1,2-dimethylhydrazine-induced colon cancer in mice by naturally occuring organosulphur compounds. Cancer Res 50:5084–5087
Lorenson MY, Cuccaro ML, Jacobs LS (1990) Calcium release from pituitary secretory granules: modulation by thiols, disulfides, and dihydropyridine calcium channel blockers. Endocrinology 126:2671–2678
Dausch JG, Nixon DW (1990) Garlic: a review of its relationship to malignant disease. Prev Med 19:346–361
Pettit GR, Hoard MS, Doubek DL, Schmidt JM, Pettit RK, Tackett LP, Chapuis JC (1996) Antineoplastic agents 338. The cancer cell growth inhibitory. Constituents of Terminalia arjuna (Combretaceae). J Ethnopharmacol 53:57–63
Neutra M, Louvard D (1989) Functional epithelial cells in culture. Alan R Liss Inc., New York, pp 363–368
Orchel A, Dzierzewicz Z, Parfiniewicz B, Weglarz L, Wilczok T (2005) Butyrate induced differentiation of colon cancer cells is PKC and JNK dependent. Dig Dis Sci 50:490–498
King J (1965). Lactate dehydrogenase. In: King J (ed) Practical clinical enzymology. D. Van Norstrand Co., London, pp 83–93
Sudhandiran G, Shaha C (2003) Antimonial-induced increase in intracellular Ca2+ through non-selective cation channels in the host and the parasite is responsible for apoptosis of intracellular Leishmania donovani amastigotes. J Biol Chem 278:25120–25132
Ho YS, Wang YJ, Lin JK (1996) Induction of p53 and p21/WAF1/CIP1 expression by nitric oxide and their association with apoptosis in human cancer cells. Mol Carcinog 16:20–31
Rasola A, Geuna M (2001) A flow cytometry assay simultaneously detects independent apoptotic parameters. Cytometry 45:151–157
Towbin H, Stahelin H, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gel to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Sundaram SG, Milner JA (1993). Impact of organosulfur components in garlic on canine mammary tumor cells in culture. Cancer Lett 74:85–90
Hosono T, Fukao T, Ogihara J, Ito Y, Shiba H, Seki T, Ariga T (2005) Diallyl trisulfide suppresses the proliferation and induces apoptosis of human colon cancer cells through oxidative modification of beta-tubulin. J Biol Chem 280:41487–41493
Yuasa H, Hirano K, Kodama H, Nakanishi H, Imai T, Tsuda H, Imaida K, Tatematsu M (1994) Immunohistochemical demonstration of intestinal-type alkaline phosphatase in stomach tumors induced by N-Methyl-N′-nitro-N-nitrosoguanidine in rats. Jpn J Cancer Res 85:897–903
Chittezhath M, Kuttan G (2006) Radioprotective activity of naturally occurring organosulfur compounds. Tumori 92:163–169
Morgan DO (1995) Principles of CDK regulation. Nature 374:131–134
Pervin S, Singh R, Freije WA, Chaudhuri G (2003) MKP-1-induced dephosphorylation of extracellular signal-regulated kinase is essential for triggering nitric oxide-induced apoptosis in human breast cancer cell lines: implications in breast cancer. Cancer Res 63:8853–8860
Cook T, Wang Z, Alber S, Liu K, Watkins SC, Vodovotz Y, Billiar TR, Blumberg D (2004) Nitric oxide and ionizing radiation synergistically promote apoptosis and growth inhibition of cancer by activating p53. Cancer Res 64:8015–8021
Trump BF, Berezesky IK, Chang SH, Phelps PC (1997) The pathways of cell death: oncosis, apoptosis, and necrosis. Toxicol Pathol 25:82–88
Goldsworthy TL, Conolly RB, Fransson-Steen R (1996) Apoptosis and cancer risk assessment. Mutat Res 365:71–90
Stuart MC, Damoiseaux JG, Frederik PM, Arends JW, Reutelingsperger CP (1998) Surface exposure of phosphatidylserine during apoptosis of rat thymocytes precedes nuclear changes. Eur J Cell Biol 76:77–83
Yang WL, Sun AY (1998) Paraquat-induced cell death in PC12 cells. Neurochem Res 23:1387–1394
Um HD, Cho YH, Kim DK, Shin JR, Lee YJ, Choi KS, Kang JM, Lee MG (2004) TNF-α suppresses dendritic cell death and the production of reactive oxygen intermediates induced by plasma withdrawal. Exp Dermatol 13:282–288
Littlewood T, Evan G (1998) A matter of life and cell death. Science 281:1317–1322
Huang R, Wallqvist A, Covell DG (2005) Anticancer metal compounds in NCI's tumor-screening database: putative mode of action. Biochem Pharmacol 69:1009–1039
Karmakar S, Banik NL, Patel SJ, Ray SK (2007) Garlic compounds induced calpain and intrinsic caspase cascade for apoptosis in human malignant neuroblastoma SH-SY5Y cells. Apoptosis 12:671–684
Kyriakis JM, Avruch J (1996) Protein kinase cascades activated by stress and inflammatory cytokines. Bioessays 18:567–577
Ip YT, Davis RJ (1998) Signal transduction by the c-Jun N-terminal kinase (JNK) from inflammation to development. Curr Opin Cell Biol 10:205–219
Impey S, Obrietan K, Storm DR (1999) Making new connections: role of ERK/MAP kinase signalling in neuronal plasticity. Neuron 23:11–14
Zelivianski S, Spellman M, Kellerman M, Kakitelashvilli V, Zhou XW, Lugo E, Lee MS, Taylor R, Davis TL, Hauke R, Lin MF (2003) ERK inhibitor PD98059 enhances docetaxel-induced apoptosis of androgen-independent human prostate cancer cells. Int J Cancer 107:478–485
Oommen S, Anto RJ, Srinivas G, Karunagaran D (2004) Allicin (from garlic) induces caspase-mediated apoptosis in cancer cells. Eur J Pharmacol 485:97–103
Acknowledgment
We wish to thank Dr. Ramamurthy, Director, Ultrafast process, University of Madras, for helping us in confocal microscopy imaging.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sriram, N., Kalayarasan, S., Ashokkumar, P. et al. Diallyl sulfide induces apoptosis in Colo 320 DM human colon cancer cells: involvement of caspase-3, NF-κB, and ERK-2. Mol Cell Biochem 311, 157–165 (2008). https://doi.org/10.1007/s11010-008-9706-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11010-008-9706-8