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The Allosteric Heat Shock Protein Inhibitor YM‐01 Protects Against Alcohol‐Induced Growth Arrest
Author(s) -
Ganapathy Ekambaram,
Khachatoorian Ronik,
Patel Harsh,
Arumugaswami Vaithilingaraja,
French Samuel
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.611.3
Subject(s) - chemistry , cyp2e1 , oxidative stress , viability assay , ethanol , lipid peroxidation , heat shock , reactive oxygen species , heat shock protein , cell growth , microbiology and biotechnology , biochemistry , apoptosis , cytochrome p450 , biology , metabolism , gene
Background Oxidative stress is implicated in the pathogenesis of alcohol‐induced liver injury. This is, in part, mediated by the cytochrome P450 CYP2E1 which contributes to reactive oxygen species (ROS) production. YM‐01, a derivative of MKT‐077, is an allosteric heat shock protein inhibitor and modulator of the triage function of heat shock cognate protein 70 (HSC70) and may protect cells from oxidative stress. In this study, we investigated the effect of YM01 on ethanol‐induced growth arrest. Methods HepG2 cells overexpressing CYP2E1 (E47) were pretreated with either vehicle or YM‐01 then treated with arachidonic acid (60 µM for 18 hrs), followed by iron nitrilotriacetic acid (Fe‐NAT, 25 µM for 5 hrs), and finally with ethanol (100 mM for 48 hrs). MTT and western analysis was then performed. Results In the CYP2E1‐expressing ethanol‐treated cells, cell growth was arrested as determined by cell viability (MTT assay). This arrest was relieved by treatment with YM‐01. Further, western analysis showed that pretreatment with YM‐01 prevented 4‐hydroxynonenal (4‐HNE) adduct formation induced by ethanol (the most abundant lipid peroxidation end product). In addition, we determined ubiquitination levels by Western analysis to determine role of the ubiquitin‐proteasome pathway. Conclusion This study indicates that YM‐01 decreases 4‐HNE adduct formation in CYP2E1‐expressing cells and protects against alcohol induced cell growth arrest.