Sulforaphane-mediated induction of a phase 2 detoxifying enzyme NAD(P)H:quinone reductase and apoptosis in human lymphoblastoid cells.
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The effect of sulforaphane on human lymphoblastoid cells originating from a patient of a high cancer risk was studied. Sulforaphane (SFN) is a naturally occurring substance of chemopreventive activity. In our study, changes in cell growth, induction of apoptosis and phase 2 enzymes as well as glutathione level were examined. Apoptosis was tested by confocal microscopy at three stages: change in mitochondrial membrane potential, caspase activation and phosphatidylserine externalization. We show that SFN increases the activity of the detoxification system: it increases quinone reductase activity at low concentration (0.5-1 μM) and raises glutathione level in a dose-dependent manner. At higher doses (2.5-10 μM) sulforaphane is a cell growth modulator, as it caused cell growth cessation (IC50 = 3.875 μM), and apoptosis inducer. The results obtained suggest that sulforaphane acts as a chemopreventive agent in human lymphoblastoid cells.
- Bonnesen C, Eggleston IM, Hayes JD. (2001) Dietary indoles and isothiocyanates that are generated from cruciferous vegetables can both stimulate apoptosis and confer protection against DNA damage in human colon cell lines. Cancer Res.; 61: 6120-30.
- Brooks JD, Paton VG, Vidanes G. (2001) Potent induction of phase 2 enzymes in human prostate cells by sulforaphane. Cancer Epidemiol Biomarkers Prev.; 10: 949-54.
- Chiao JW, Chung FL, Kancherla R, Ahmed T, Mittelman A, Conaway DD. (2002) Sulforaphane and its metabolite mediate growth arrest and apoptosis in human prostate cancer cells. Int J Oncol.; 20: 631-6.
- Clarke RG, Lund EK, Johnson IT, Pinder AC. (2002) Apoptosis can be detected in attached colonic adenocarcinoma HT29 cells using annexin V binding, but not by TUNEL assay or sub-G0 DNA content. Cytometry.; 39: 141-50.
- Fimognari C, Nusse M, Berti F, Iori R, Cantelli-Forti G, Hrelia P. (2002) Cyclin D3 and p53 mediate sulforaphane-induced cell cycle delay and apoptosis in non-transformed human T lymphocytes. Cell Mol Life Sci.; 59: 2004-12.
- Fimognari C, Nüsse M, Cesari R, Iori R, Cantelli-Forti G, Hrelia P. (2002) Growth inhibition, cell cycle arrest and apoptosis in human T-cell leukemia by the isothiocyanate sulforaphane. Carcinogenesis.; 23: 581-6.
- Gamet-Payrastre L, Lumeau S, Gasc N, Cassar G, Rollin P, Tulliez J. (1998) Selective cytostatic and cytotoxic effects of glucosinolates hydrolysis products on human colon cancer cells in vitro. Anticancer Drugs.; 9: 141-8.
- Gamet-Payrastre L, Li P, Lumeau S, Cassar G, Dupont MA, Chevolleau S, Gasc N, Tulliez J, Terce F. (2000) Sulforaphane, a naturally occurring isothiocyanate, induces cell cycle arrest and apoptosis in HT29 human colon cancer cells. Cancer Res.; 60: 1426-33.
- Gamet-Payrastre L, Li P, Lumeau S, Cassar G, Dupont MA, Chevolleau S, Gase N, Tulliez J, Terce F. (2000) Sulforaphane, a naturally occurring isotiocyanate, induces cell cycle arrest and apoptosis in HT29 human colon cancer cells. Cancer Res.; 60: 1426-33.
- Goerke A, Sakai N, Gutjahr E, Shlapkohl WA, Mushinski JF, Haller H, Kolch W, Saito N, Mischak H. (2002) Induction of apoptosis by protein kinase C is independent of its kinase activity. J Biol Chem.; 277: 32054-62.
- Hedley D, Chow S. (1994) Glutathione and cellular resistance to anti-cancer drugs. Methods Cell Biol.; 42: 31-44.
- Hug H, Los M, Hirt W, Debatin KM. (1999) Rhodamine 110-linked amino acids and peptides as substrates to measure caspase activity upon apoptosis induction in intact cells. Biochemistry.; 38: 13906-11.
- Iori R, Bernardi D, Gueyrard P, Rollin S, Palmieri S. (1999) Formation of glucoraphanin by chemoselective oxidation of natural glucoerucin: a chemoenzymatic route to sulforaphane. Bioorg Med Chem Lett.; 9: 1047-8.
- Jiang ZQ, Chen C, Yang B, Hebbar V, Kong AN. (2003) Differential responses from seven mammalian cell lines to the treatments of detoxifying enzyme inducers. Life Sci.; 72: 2243-53
- Kolm RH, Danielson UH, Zhang Y, Talalay P, Mannervik B. (1995) Isothiocyanates as substrates for human glutathione transferases: structure-activity studies. Biochem J.; 311: 453-9.
- Leoni O, Iori R, Palmieri S. (2000) Hydrolysis of glucosinolates using nylon-immobilized myrosinase to produce pure bioactive molecules. Biotechnol Bioeng.; 68: 660-4.
- Misiewicz I, Skupinska K, Kasprzycka-Guttman T. (2003) Sulforaphane and 2-oxoheksyl isothiocyanate induce cell growth arrest and apoptosis in L-1210 leukemia and ME-18 melanoma cells. Oncol Rep.; 10: 2045-50.
- Palmer AM, Greengrass PM, Cavalla D. (2000) The role of mitochondria in apoptosis. Drug News Perspect.; 13: 378-84.
- Pessina A, Thomas RM, Palmieri S, Luisi PL. (1990) An improved method for the purification of myrosinase and its physicochemical characterization. Arch Biochem Biophys.; 280: 383-9.
- Prohaska HJ, Santamaria AB. (1988) Direct measurement of NAD(P)H:quinone reductase from cells cultured in microtiter wells: a screening assay for anticarcinogenic enzyme inducers. Anal Biochem.; 169: 328-36.
- Purring-Koch C, McLendon G. (2000) Cytochrome c binding to Apaf-1: The effects of dATP and ionic strength. Proc Natl Acad Sci USA.; 97: 11928-31.
- Rosa EAS, Heaney RK, Fenwick GR, Portas CAM. (1997) Glucosinolates in crop plants. Horticult Rev.; 19: 99-215.
- Schmidt H, Karrer P. (1948) Synthese der Racemischen und der Optisch Aktiven Formen des Sulforaphans. Hel Chim Acta.; 31: 1497-505.
- Seamus JM, Finucane DM, Amarante-Mendez GP, O'Brien GA, Green DR. (1996) Phosphatidylserine externalization during CD95-induced apoptosis of cells and cytoplasts requires ICE/CED-3 protease activity. J Biol Chem.; 271: 28753-6.
- Smiley ST, Reers M, Mottola-Hartshorn C, Lin M, Chen A, Smith TW, Steele GD Jr, Chen LB. (1991) Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate-forming lipophilic cation JC-1. Proc Natl Acad Sci USA.; 88: 3671-5.
- Steinkeller H, Rabot S, Freyald C, Nobis E, Scharf G, Chabicovsky M, Knasmuller S, Kassie F. (2001) Effect of cruciferous vegetable and their constituents on drug metabolizing enzymes involved in the bioactivation of DNA-reactive dietary carcinogens. Mutat Res.; 480/481: 285-97.
- Tawfiq N, Heaney RK, Plumb JA, Fenwick GA, Musk SR, Williamson G. (1995) Dietary glucosinolates as blocking agents against carcinogenesis: glucosinolate breakdown products assessed by induction of quinone reductase activity in murine hepa1c1c7 cells. Carcinogenesis.; 16: 1191-4.
- Tietze F. (1969) Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal Biochem.; 27: 502-22.
- Wattenberg LW. (1990) Inhibition of carcinogenesis by minor anutrient constituents of the diet. Proc Nutr Soc.; 49: 173-83.
- Willingham MC. (1999) Cytochemical methods for the detection of apoptosis. J Histochem Cytochem.; 47: 1101-9.
- Ye L, Dinkova-Kostova AT, Wade KL, Zhang Y, Shapiro TA, Talalay P. (2002) Quantitative determination of dithiocarbamates in human plasma, serum, erythrocytes and urine: pharmacokinetics of broccoli sprouts isothiocyanates in humans. Clin Chim Acta.;316: 43-53.
- Yu R, Mandlekar S, Harvey KJ, Ucker DS, Kong AN. (1998) Chemopreventive isothiocyanates induce apoptosis and caspase-3-like protease activity. Cancer Res.; 58: 402-8.
- Zhang Y, Talalay P. (1998) Mechanism of differential potencies of isothiocyanates as inducers of anticarcinogenic phase 2 enzymes. Cancer Res.; 58: 4632-9.
- Zhang Y, Callaway EC. (2002) High cellular accumulation of sulphoraphane, a dietary anticarcinogen, is followed by rapid transporter-mediated export as a glutathione conjugate. Biochem J.; 364: 301-7.
- Zhang Y, Talalay P, Cho C, Posner GA. (1992) A major inducer of anticarcinogenic protective enzymes from broccoli: isolation and elucidation of structure. Proc Natl Acad Sci USA.; 89: 2399-403.
- Zhao Q, Yang LX, Holtzlaw WD, Talalay P. (1997) Unexpected genetic and structural relationships of a long-forgotten flavoenzyme to NAD(P)H:quinone reductase (DT-diaphorase). Proc Natl Acad Sci USA.; 94: 1669-74.
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