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

Phoebe Trio,Satoru Fujisaki,De-Xing Hou,Atsuyoshi Kawahara

doi:10.26534/kimika.v29i1.1-10

Abstract

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.

Highlights

Isothiocyanate (ITC) is a small but reactive organosulfur containing molecule present in most cruciferous vegetables
Comparison of the direction of regulation showed that 6-(methylsulfinyl)hexyl isothiocyanate (6-MSITC) and 6-(methylthio)hexyl isothiocyanate (6-MTITC) had stronger effect towards the downregulation of IMR-32 genes
It should be noted that the reported studies evaluated structure-bioactivity relationship of different kinds of ITC and not the genome-wide expression effect. This is vital since this is the first report to demonstrate global gene expression of changes induced by SFN and SFN analogues and this inconsistency could be possibly attributed to the cell variation which is in accordance with the results of Trio et al, (2017) and Trio et al, (2016)

Summary

Introduction

Isothiocyanate (ITC) is a small but reactive organosulfur containing molecule present in most cruciferous vegetables. This study was designed to compare the global changes in transcript levels and the underlying genes targeted by Wasabi-derived ITCs in hepatic cells with that of neuronal cells using microarray-based technology. Microarray-based gene transcription profiling to contrast the genome wide-gene expression changes associated with Wasabi SFN, 6-MSITC and 6-MTITC stimulation in HepG2 cells with that of IMR-32 cells. The previous studies indicated that 6-MTITC is the strongest inducer of gene expression changes in HepG2 cells while 6-MSITC is the strongest inducers in IMR-32 cells.

Results

Conclusion