Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/15618
Title: A proteomic approach to study the apoptotic effects of phenethyl isothiocyanate on the human hepatoma cell line, HepG2
Authors: NEO CHUN HONG, JASON
Keywords: phenethyl isothiocyanate, proteomics, mass spectrometry, difference gel electrophoresis, chemoprevention, MALDI
Issue Date: 8-Nov-2006
Source: NEO CHUN HONG, JASON (2006-11-08). A proteomic approach to study the apoptotic effects of phenethyl isothiocyanate on the human hepatoma cell line, HepG2. ScholarBank@NUS Repository.
Abstract: B-phenylethyl isothiocyanate (PEITC) is a promising chemopreventative agent found abundance in watercress (Rorripa nasturtium aquaticum) as its glucosinolate precursor. In the present investigation, we sought to determine the early changes in protein expression that contribute to the mechanism(s) of PEITC mediated apoptosis in the human hepatoma HepG2 cell line. Such data may lead to the identification of new molecular targets of PEITC, this contributing to a greater understanding of the mechanism(s) by which isothiocyanates (ITCs) mediate apoptotic cascades. Using two dimensional difference gel electrophoresis we determined the changes in global protein expression between control (0.01% DMSO) and PEITC (IC50~ 20A?M) treated cells after 3- and 6-h, such time points being chosen to circumvent the effects of caspase mediate proteolysis. Comparison between PEITC treated cells with their respective controls showed that seventeen protein spots were differentially expressed. Fourteen of these spots, representing nine unique proteins, were successfully identified using MALDI-TOF and MALDI TOF/TOF mass spectrometry. We observed significant shifts in pIs on 2-DE gels in heat shock 27 kDa protein (Hsp27), macrophage migration inhibition factor (MIF) and heterogeneous nuclear ribonucleoprotein K (hnRNP K) indicating that these proteins are probably involved in protein phosphorylation. Indeed, hnRNP K was determined to be phosphorylated on key tyrosine residues as assessed by anti-phosphotyrosine antibodies. In separate experiments we also showed that c-myc is up-regulated in PEITC treated cells, and since hnRNP K is reported to induce over-expression of c-myc, we proposed that PEITC-induced apoptosis may involve a c-myc dependent apoptotic pathway in HepG2 cells.
URI: http://scholarbank.nus.edu.sg/handle/10635/15618
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01_Introduction.PDF82.75 kBAdobe PDF

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02_Materials_and_Methods.PDF104.76 kBAdobe PDF

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03_Results.PDF5.49 MBAdobe PDF

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04_Discussion.PDF58.8 kBAdobe PDF

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05_Conclusion.PDF27.22 kBAdobe PDF

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06_References.PDF53.88 kBAdobe PDF

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07_List of Publications.PDF12.93 kBAdobe PDF

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08_Appendix.PDF3.57 MBAdobe PDF

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