Comparative Gene Expression Profiling in Cultured Cell Lines after Treatment with Wasabi-derived Isothiocyanates

  • Phoebe Trio Chemistry Department, College of Science, De La Salle University – Manila
  • Atsuyoshi Kawahara Department of Biochemical Sciences and Biotechnology, Faculty of Agriculture, Kagoshima University, Korimoto 1-21-24, Kagoshima 890-0065
  • Satoru Fujisaki Department of Biochemical Sciences and Biotechnology, Faculty of Agriculture, Kagoshima University, Korimoto 1-21-24, Kagoshima 890-0065
  • De-Xing Hou Department of Biochemical Sciences and Biotechnology, Faculty of Agriculture, Kagoshima University, Korimoto 1-21-24, Kagoshima 890-0065; United Graduate School of Agricultural Sciences, Kagoshima University, Korimoto 1-21-24, Kagoshima 890-0065
Keywords: gene expression profiling, isothiocyanates, Japanese wasabi


Isothiocyanate (ITC) is a small but reactive organo-sulfur containing group of compound present in cruciferous vegetables. Compendium of evidence indicated that ITCs exhibit multiple biological activities, but the exact molecular mechanisms are not yet clear. Therefore, this study was designed to compare the genome-wide gene expression profiles in hepatic and neuron cells following wasabi-derived ITCs treatment using microarray technology. Cells were treated with wasabi-derived ITCs, sulforaphane (SFN), 6-(methylsulfinyl)hexyl isothiocyanate (6-MSITC) and 6-(methylthio)hexyl isothiocyanate (6-MTITC), for 9 h and was followed by DNA microarray analyses using HG-U133 plus 2.0 oligonucleotide array. Selected gene products were confirmed by real-time PCR, and functional subsets of genes and biologically significant network were identified using Ingenuity Pathway Analysis. Results showed that 6-MTITC was the most potent inducer of gene expressions changes in HepG2 cells, whereas 6-MSITC was the most effective inducer in IMR-32 cells. Despite this cell-type response discrepancies, 6-MSITC came up as the strongest inducer of antioxidant-associated genes, via the regulation of the Nrf2-mediated pathway. These results combined with the varying induction level data of other Wasabi-derived ITCs form the basis for further studies to assess the possible therapeutic effect of combined Wasabi-derived ITCs treatment. Altogether, this study provided comprehensive information on how structural differences of Wasabi-derived ITCs contribute to its efficacy and impact specific targets.


Adesina AM, Lopez-Terrada D, Wong KK, Gunaratne P, Ngyuyen Y, Pulliam J, Margolin J, and Finegold M: Gene expression profiling reveals signatures characterizing histologic subtypes of hepatoblastoma and global deregulation in cell growth and survival pathways. Hum Pathol 40, 843–853, 2009.
Atwell LL, Beaver LM, Shannon J, Williams DE, Dashwood RH, and Ho E: Epigenetic regulation by sulforaphane: Opportunities for breast and prostate cancer chemoprevention. Curr Pharmacol Rep 1, 102–111, 2015.
Babish JG, Pacioretty LM, Bland JS, Minich DM, Hu J, and Tripp ML: Antidiabetic screening of commercial botanical products in 3T3-L1 adipocytes and db/db mice. J Med Food 13, 535–547, 2010.
Chaudhuri D, Orsulic S, and Ashok BT: Antiproliferative activity of sulforaphane in Akt-overexpressing ovarian cancer cells. Mol Cancer Ther 6, 334–345, 2007.
Chen J, Uto T, Tanigawa S, Yamada-Kato T, Fujii M, and Hou DX: Microarray-based determination of anti-inflammatory genes targeted by 6-(methylsulfinyl)hexyl isothiocyanate in macrophages. Exp Ther Med 1, 33–40, 2010.
Chung YK, Or RCH, Lu CH, Ouyang WT, Yang SY, and Chang CC: Sulforaphane down-regulates SKP2 to stabilize p27KIP1 for inducing anti-proliferation in human colon adenocarcinoma cells. J Biosci Bioeng 119, 35–42, 2015.
Hou DX, Fukuda M, Fujii M, and Fuke Y: Transcriptional regulation of nicotinamide adenine dinucleotide phosphate: quinone oxidoreductase in murine hepatoma cells by 6-(methylsulfinyl)hexyl isothiocyanate, an active principle of wasabi (Eutrema wasabi Maxim.). Cancer Lett 20, 195–200, 2000.
Isshiki K and Tokouka K: Allyl Isothiocyanate and wholesomeness of food. Jpn J Food Microbiol 12, 1–6, 1993.
Itoh K, Chiba T, Takahashi S, Ishii T, Igarashi K, Katoh Y, Oyake T, Hayashi N, Satoh K, Hatayama I, Yamamoto M, and Nabeshima Y: An Nrf2/small Maf heterodimer mediates the induction pf phase II detoxifying enzyme genes through antioxidant response element. Biochem Biophys Res Commun 236, 313–322, 1997.
Keiko F, and Sasahara M: The roles of PDGF in development and during neurogenesis in the normal and diseased nervous system. J Neuroimmune Pharmacol, 9, 168–181, 2014.
Kim YJ, Lee DH, Ahn J, Chung WJ, Kang YJ Seong KS, Moon JH, Ha TY, and Jung CH: Pharmacokinetics, tissue distribution, and anti-lipogennic/adipogenic effects of allyl isothiocyanates metabolites. PLOS ONE 10, e0132151, 2015.
Kumagai H, Kashima N, Seki T, Sakurai H, Ishii K, and Ariga T: Analysis of components in essential oil of upland Wasabi and their inhibitory effects on platelet aggregation. Biosci Biotech Biochem 58, 2131–2135, 1994.
Lahiri DK: The stability of beta-amyloid precursor protein in nine different cell types. Biochem Mol Biol Int 29, 849–858, 1993.
Lee YS: Anti-oxidant effect of Wasabi Japonica extracts. Korean J Oriental Prev Medical Soc 12, 119–126, 2008.
Lemerchant S, Pruvost M, Montaner J, Emery E, Vivien D, Kanninen K, and Koistinaho J: ADAMTS proteoglycanases in the physiological and pathological central nervous system. J Neuroinflammation 10, 133–140, 2013.
Lopez-Terrada D, Gunaratne PH, Adesina AM, Pulliam J, Hoang DM, Nguyen Y, Mistretta TA, Margolin J, and Finegold MJ: Histologic subtypes of hepatoblastoma are characterized by the differential canonical Wnt and Notch patway activation in DLK+ precursors. Hum Path 40, 783–794, 2009.
Mazzarello P: A unifying concept: the history of cell theory. Nat Cell Bio 1, E13–E15, 1999.
Moehlenkamp J, and Johnson JA: Activation of antioxidant/electrophile-responsive elements in IMR-32 human neuroblastoma cells. Arch Biochem Biophys 363, 98–106, 1999.
Myzak MC, Dashwood WM, Orner GA, Ho E, and Dashwood RH: Sulforaphane inhibits histone deacetylase in vivo and suppresses tumorigenesis in Apcmin mice. FASEB J 20, 506–508, 2006.
Neerven SV, Kampmann E, and Mey J: RAR/RXR and PPAR/RXR signaling in neurological and psychiatric diseases. Prog Neurobiol 85, 433¬–451, 2008.
Nguyen T, Huang HC, and Pickett CB: Transcriptional regulation of the antioxidant response element: Activation by Nrf2 and repression by MafK. J Biol Chem 275, 15466–15473, 2000.
Okamoto T, Akita N, Nagai M, Hayashi T, and Suzuki K: 6-Methylsulfinylhexyl isothiocyanate modulates endothelial cell function and suppresses leukocyte adhesion. J Nat Med 68, 144–153, 2013.
Ono H, Tesaki S, Tanabe S, and Watanabe M: 6-Methylsulfinylhexyl isothiocyanate and its homologues as food-originated compounds with antibacterial activity against Escherichia coli and Staphylococcus aureus. Biosci Biotechnol Biochem 62, 363–365, 1998.
Ryu HY, Kyung HB, Eun JK, Sang JP, Bong L, and HO YS: Evaluation for the antimicrobial, antioxidant and antithrombosis activity of natural spices for fresh cut yam. J Life Sci 17, 652–657, 2007.
Shin IS, Masuda H, and Naohide K: Bactericidal activity of wasabi (Wasabia japonica) against Helicobacter pylori. Int J Food Microbiol 94, 255–261, 2004.
Shin SW, Ghimeray AK, and Park CH: Investigation of total phenolic, total flavonoid, antioxidant and allyl isothiocyanate content in the different organs of Wasabi japonica grown in an organic system. Afr J Tradit Complement Altern Med 11, 38–45, 2014.
Sidransky H, Ito N, and Verney E: Influence of alpha-naphthyl-isothiocyanate on liver tumorigenesis in rats ingesting ethionine and N-2-flourenylacetamide. J Natl Cancer Inst 37, 677–686, 1966.

Sun CC, Li SJ, Yang CL, Xue RL, Xi YY, Wang L, Zhao QL, and Li DJ: Sulforaphane attenuates muscle inflammation in dystrophin-deficient mdx mice via NF-E2-related factor 2 (Nrf2)-mediated inhibition of NF-κB signaling pathway. J Biol Chem 290, 17784–17795, 2015.
Tarozzi A, Angeloni C, Malaguti M, Morroni F, Hrelia S, and Hrelia P: Sulforaphane as a potential protective phytochemical against neurodegenerative diseases. Oxid Med Cell Longev 2013, 415078, 2013. 

Trio PZ, Fujisaki S, Tanigawa S, Hisanaga A, Sakao K, Hou DX: DNA microarray highlights Nrf2-mediated neuron protection targeted by wasabi-derived isothiocyanates. Gene Regul Sys Bio 10, 73–83, 2016.
Trio PZ, Kawahara A, Tanigawa S, Sakao K, Hou DX: DNA microarray profiling highlights Nrf2-mediated chemoprevention taegeted by Wasabi-derived isothiocyanates in HepG2 cells. Nutr Cancer 69, 105–116, 2017.
Uto T, Fujii M, and Hou DX: Inhibition of lipopolysaccharide–induced cyclooxygenase-2 transcription by 6-(methylsulfinyl) hexyl isothiocyanate, a chemopreventive compound from Wasabia japonical (Miq.) Matsumura, in mouse macrophages. Biochem Pharmacol 70, 1772–1784, 2005a.
Uto T, Fujii M, and Hou DX: 6-(Methylsulfinyl)hexyl isothiocyanate suppresses inducible nitric oxide synthase expression through the inhibition of Janus kinase 2-mediated JNK pathway in lipopolysaccharide-activated murine macrophage. Biochem Pharmacol 70, 1211–1221, 2005b.
Uto T, Fujii M, and Hou DX: Effects of 6-(methylsulfinyl)hexyl isothiocyanate on cyclooxygenase-2 expression induced by lipopolysaccharide, interferon-γ and 12-O-tetradecanoylphorbol-13-acetate. Oncol Rep 17, 233–238, 2007.
Valledor AF, Hsu LC, Ogawa S, Sawka-Verhelle D, Karin M, and Glass CK: Activation of liver X receptors and retinoid X receptors prevents bacterial-induced macrophage apoptosis. PNAS 101, 17813–17818, 2004.
Vasanthi HR, Mukherjee S, and Das DK: Health benefits of broccoli- a chemico-biological overview. Min Rev Med Chem 9, 749–759, 2009.
Ye L, Yu T, Li Y, Chen B, Zhang J, Wen Z, Zhang B, Zhou X, Li X, Li F, Cao W, and Huang Z: Sulforaphane enhances the ability of human retinal pigment epithelial cell against oxidative stress, and its effect on gene expression profile evaluated by microarray analysis. Oxid Med Cell Longev 2013, 413024, 2013. 

Zhang Y, Talalay P, Cho CG, and Posner GH: A major inducer of anticarcinogenic protective enzymes from broccoli: isolation and elucidation of structure. Proc Natl Acad Sci U S A 89, 2399–2403, 1992.
How to Cite
Trio, P., Kawahara, A., Fujisaki, S., & Hou, D.-X. (2018). Comparative Gene Expression Profiling in Cultured Cell Lines after Treatment with Wasabi-derived Isothiocyanates. KIMIKA, 29(1), 1-10.
Research Articles