Allyl Isothiocyanate and Its N-Acetylcysteine Conjugate Suppress Metastasis via Inhibition of Invasion,Migration,and Matrix Metalloproteinase-2/-9 Activities in SK-Hep1 Human Hepatoma Cells
Restricted accessResearch articleFirst published online April, 2006
Allyl Isothiocyanate and Its N-Acetylcysteine Conjugate Suppress Metastasis via Inhibition of Invasion,Migration,and Matrix Metalloproteinase-2/-9 Activities in SK-Hep1 Human Hepatoma Cells
Cruciferous vegetables contain a series of relatively unique secondary metabolites of amino acids, called glucosinolates. Sinigrin, the predominant aliphatic glucosinolate in cruciferous vegetables, is hydrolyzed to yield allyl isothiocyanate (AITC), which, after absorption and metabolism in humans, is excreted in the urine as an N-acetyicysteine (NAC) conjugate. We have determined the inhibitory effects of AITC and its NAC conjugate on cell proliferation, the expression of matrix metalloproteinases (MMPs), adhesion, invasion, and migration in SK-Hep1 human hepatoma cells. Our results demonstrate that AITC and NAC-AITC suppress SK-Hep1 cell proliferation in a dose-dependent manner; by 25% and 30% for 10 μM AITC and 10 μM NAC-AITC, respectively. We examined the influence of AITC and NAC-AITC on the gene expression of MMPs and tissue inhibitors of metalloproteinase (TIMPs). Gelatin zymography also revealed a significant downregulation of MMP-2/-9 expression in SK-Hep1 cells treated with 0.1–5 μM AITC and NAC-AITC compared with controls. Reverse transcriptase polymerase chain reaction revealed dose-dependent decreases in MMP-2/-9 messenger RNA levels in both AITC-treated and NAC-AITC–treated cells. TIMP-1/-2 activities were unaffected by treatment with AITC or NAC-AITC in our experiments. NAC-AITC inhibited cancer cell adhesion and invasion much more potently than its parent compound. NAC-AITC at 5 μM caused excellent inhibition of cell migration for 48 hrs. These results demonstrate the potential of AITC and NAC-AITC as chemopreventive agents.
LampeJW, PetersonS. Brassica, biotransformation and cancer risk: genetic polymorphisms alter the preventive effects of cruciferous vegetables. J Nutr132: 2991–2994, 2002.
2.
FenwickGR, HeaneyRK. Glucosinolates and their breakdown products in cruciferous crops, foods and feeding stuffs. Food Chem11: 249–271, 1983.
3.
MennickeWH, KralT, KrumbiegelG. Metabolism of some naturally occurring isothiocyanates in the rat. Xenobiotica13: 203–207, 1983.
4.
HwangE, JefferyEH. Evaluation of urinary N-acetyl cysteinyl allyl isothiocyanate as a biomarker for intake and bioactivity of Brussels sprouts. Food Chem Toxicol41: 1817–1825, 2003.
5.
MundayR, MundayCM. Selective induction of phase II enzymes in the urinary bladder of rats by allyl isothiocyanate, a compound derived from Brassica vegetables. Nutr Cancer44: 52–59, 2002.
6.
HwangE, LeeHJ. Induction of quinine reductase by allylisothiocyanate (AITC) and the N-acetylcysteine conjugate of AITC in Hepa1c1c7 mouse hepatoma cells. BioFactors (in press), 2006.
7.
ManeshC, KuttanG. Effect of naturally occurring allyl and phenyl isothiocyanates in the inhibition of experimental pulmonary metastasis induced by B16F-10 melanoma cells. Fitoterapia74: 355–363, 2003.
8.
ConawayCC, YangYM, ChungF-L. Isothiocyanates as cancer chemopreventive agents: their biological activities and metabolism in rodents and humans. Curr Drug Metab3: 233–255, 2002.
9.
ChiaoJW, ChungF-L, RancherlaR, AhmedT, MittelmanA, ConawayCC. Sulforaphane and its metabolite mediate growth arrest and apoptosis in human prostate cancer cells. Int J Oncol20631–636, 2002.
10.
ChiaoJW, ChungF-L, KrzeminskiJ, AminS, AshadR, AhmedT, ConawayCC. Modulation of growth of prostate cancer cells by the N-acetylcysteine conjugate of phenethyl isothiocyanate. Int J Oncol16: 1215–1219, 2000.
11.
ChiaoJW, WuH, RamaswamyG, ConawayCC, ChungF-L, WangL, LiuD. Ingestion of an isothiocyanate metabolite from cruciferous vegetables inhibits growth of human prostate cancer cell xenografts by apoptosis and cell cycle arrest. Carcinogenesis25: 1403–1408, 2004.
12.
HwangE, JefferyEH. Induction of quinine reductase by sulforaphane and sulforaphane N-acetylcysteine conjugate in murine hepatoma cells. J Med Food8: 198–203, 2005.
13.
HechtSS, UpadhyayaP, WangM, BlissRL, MclnteeEJ, KenneyPM. Inhibition of lung tumorigenesis in A/J mice by N-acetyl-S-(N-2-phenethylthiocarbamoyl)-L-cysteine and myo-inositol, individually and in combination. Carcinogenesis23: 1455–1461, 2002.
14.
PriceJT, ThompsonEW. Mechanisms of tumour invasion and metastasis: emerging targets for therapy. Expert Opin Ther Targets6: 217–233, 2002.
15.
Simpson-HaidarisPJ, RybarczykB. Tumors and fibrinogen. The role of fibrinogen as an extracellular matrix protein. Ann N Y Acad Sci936: 406–425, 2001.
16.
KleinG, VellengaE, FraaijeMW, KampsWA, de BontES. The possible role of matrix metalloproteinase (MMP)-2 and MMP-9 in cancer, e.g. acute leukemia. Crit Rev Oncol Hematol50: 87–100, 2004.
17.
ZuckerS, VacircaJ. Role of matrix metalloproteinases (MMPs) in colorectal cancer. Cancer Metastasis Rev23: 101–117, 2004.
18.
VincentMP. The matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) genes. Transcriptional and posttranscriptional regulation, signal transduction and cell-type-specific expression. Methods Mol Biol151: 121–148, 2001.
19.
CombourieuB, ElfolL, DelortA-M, RabotS. Identification of new derivatives of sinigrin and glucotropaeolin produced by the human digestive microflora using 1H NMR spectroscopy analysis of in vitro incubations. Drug Metab Dispos29: 1440–1445, 2001.
20.
ChambersAF, MatrisianLM. Changing views of the role of matrix metalloproteinases in metastasis. J Natl Cancer Inst89: 1260–1383, 1997.
21.
HojillaCV, MohammedFF, KhokhaR. Matrix metalloproteinases and their tissue inhibitors direct cell fate during cancer development. Br J Cancer17: 1817–1821, 2003.
22.
GomezDE, AlonsoDF, YoshijiH, ThorgeirssonUP. Tissue inhibitors of metalloproteinases: structure, regulation and biological functions. Eur J Cell Biol74: 111–122, 1997.
23.
PowellWC, KnoxJD, NavreM, GroganTM, KittelsonJ, NagleRB, BowdenGT. Expression of the metalloproteinase matrilysin in DU-145 cells increases their invasive potential in severe combined immunodeficient mice. Cancer Res53: 417–422, 1993.
24.
LokeshwarBL, SelzerMG, ZhuBQ, BlockNL, GolubLM. Inhibition of cell proliferation, invasion, tumor growth and metastasis by an oral non-antimicrobial tetracycline analog (COL-3) in a metastatic prostate cancer model. Int J Cancer98: 297–309, 2002.
25.
CaoJ, ChiarelliC, KozarekarP, AdlerHL. Membrane type 1-matrix metalloproteinase promotes human prostate cancer invasion and metastasis. Thromb Haemost93: 770–778, 2005.
26.
SehgallI, ForbesK, WebbMA. Reduced secretion of MMPs, plasminogen activators and TIMPS from prostate cancer cells derived by repeated metastasis. Anticancer Res23: 39–42, 2003.
27.
LichtinghagenR, MusholtPB, LeinM, RomerA, RudolphB, KristiansenG, HauptmannS, SchnorrD, LoeningSA, JungK. Different mRNA and protein expression of matrix metalloproteinases 2 and 9 and tissue inhibitor of metalloproteinases 1 in benign and malignant prostate tissue. Eur Urol42: 398–406, 2002.
28.
RiddickAC, ShuklaCJ, PenningtonCJ, BassR, NuttallRK, HoganA, SethiaKK, EllisV, CollinsAT, MaitlandNJ, BallRY, EdwardsDR. Identification of degradome components associated with prostate cancer progression by expression analysis of human prostatic tissues. Br J Cancer92: 2171–2180, 2005.
29.
WilsonMJ, SellersRG, WiehrC, MelamudO, PeiD, PeehlDM. Expression of matrix metalloproteinase-2 and -9 and their inhibitors, tissue inhibitor of metalloproteinase-1 and -2, in primary cultures of human prostatic stromal and epithelial cells. J Cell Physiol191: 208–216, 2002.
30.
LiuXH, RoseDP. Suppression of type IV collagenase in MDA-MB-435 human breast cancer cells by eicosapentaenoic acid in vitro and in vivo. Cancer Lett92: 21–26, 1995.
31.
RouzaudG, YoungSA, DuncanAJ. Hydrolysis of glucosinolates to isothiocyanates after ingestion of raw or microwaved cabbage by human volunteers. Cancer Epidemiol Biomarkers Prev13: 125–131, 2004.
32.
TsengE, Scott-RamsayEA, MorrisME. Dietary organic isothiocyanates are cytotoxic in human breast cancer MCF-7 and mammary epithelial MCF-12A cell lines. Exp Biol Med229: 835–842, 2004.
33.
ParnaudG, LiP, CassarG, RouimiP, TulliezJ, CombaretL, Gamet-PayrastreL. Mechanism of sulforaphane-induced cell cycle arrest and apoptosis in human colon cancer cells. Nutr Cancer48: 198–206, 2004.
34.
XiaoD, ChoiS, LeeYJ, SinghSV. Role of mitogen-activated protein kinases in phenethyl isothiocyanate-induced apoptosis in human prostate cancer cells. Mol Carcinog43: 130–140, 2005.
35.
HuangSC, HoCT, Lin-ShiauSY, LinJK. Carnosol inhibits the invasion of B16/F10 mouse melanoma cells by suppressing metalloproteinase-9 through down-regulating nuclear factor-kappa B and c-Jun. Biochem Pharmacol69: 221–232, 2005.
36.
RudekMA, VenitzJ, FiggWD. Matrix metalloproteinase inhibitors: do they have a place in anticancer therapy?Pharmacotherapy22: 705–720, 2002.
37.
MannelloF, TontiG, PapaS. Matrix metalloproteinase inhibitors as anticancer therapeutics. Curr Cancer Drug Targets5: 285–298, 2005.
38.
RatzingerG, StoitznerP, EbnerS, LutzMB, LaytonGT, RainerC, SeniorRM, ShipleyJM, FritschP, SchulerG, RomaniN. Matrix metalloproteinases 9 and 2 are necessary for the migration of Langerhans cells and dermal dentritic cells from human and murine skin. J Immunol168: 4361–4371, 2002.
39.
RoseP, HuangQ, OngCN, WhitemanM. Broccoli and watercress suppress matrix metalloproteinase-9 activity and invasiveness of human MDA-MA-231 breast cancer cells. Toxicol Appli Pharm209: 105–113, 2005.
40.
LeeJ-Y, MoonS-K, HwangC-W, NamK-S, KimY-K. Yoon H-D, Kim M-G, Kim C-H. A novel function of benzyl isothiocyanate in vascular smooth muscle cells: the role of ERK1/2, cell cycle regulation, and matrix metalloproteinase-9. J. Cell Physiol203: 493–500, 2005.
41.
NakatsukasaH, AshidaK, HigashiT, OhguchiS, TsuboiS, HinoN, NousoK, UrabeY, KinugasaN, YoshidaK, UematsuS, IshizakiM, KobayashiY, TsujiT. Cellular distribution of transcripts for tissue inhibitor of metalloproteinases 1 and 2 in human hepatocellular carcinomas. Hepatology24: 82–88, 1996.
42.
KrugerA, ArltMJ, GergM, KopitzC, BernardMM, ChangM, MobasheryS, FridmanR. Antimetastatic activity of a novel mechanism-based gelatinase inhibitor. Cancer Res65: 3523–3526, 2005.
43.
BramhallSR, StampGW, DunnJ, LemoineNR, NeoptolemosJP. Expression of collagenase (MMP-2), stromelysin (MMP-3) and tissue inhibitor of the metalloproteinases (TIMP-1) in pancreatic and ampullary disease. Br J Cancer73: 972–978, 1996.
44.
WoodM, FudgeK, MohlerJL, FrostAR, GarciaF, WangM, StearnsME. In situ hybridization studies of metalloproteinase 2 and 9 and TIMP-I and TIMP-2 expression in human prostate cancer. Clin Exp Metastasis15: 246–258, 1997.
45.
DongZ, NemethZA, CherML, PalmerKC, BrightRC, FridmanR. Differential regulation of matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 expression in co-cultures of prostate cancer and stromal cells. Int J Cancer93: 507–515, 2001.
