title: ETS2 regulating neurodegenerative signaling pathway of human neuronal (SH-SY5Y) cells exposed to single and repeated low-dose sarin (GB).
abstract: The mechanistic understanding of low-level sarin-induced neurotoxicity after single or repeated doses has yet to be explored at a cellular level. Using the microarray (Affymetrix-GeneChips) transcription profiling approach, the present study examined gene expression in human SH-SY5Y cells exposed to single (3 and 24 h) or repeated (2 x 24 h) doses of sarin (5 microg/mL) to delineate the possible mechanism. Two hundred twenty-four genes whose expression was significantly (P < 0.01) altered by at least 3-fold were selected by GeneSpringGX analysis. The comparative gene expression data confirmed the transcriptional changes to be related to dose and exposure time of sarin. The effect of a single noncytotoxic sarin dose on gene transcription was variable, whereas repeated doses over 48 h persistently down-regulated genes linked to neurodegenerative mechanisms. Thirty persistently altered genes were validated using real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Similar qRT-PCR profiles obtained in sarin-treated SH-SY5Y and HCN-1A cells confirmed the cell-independent alterations in expression levels. Genes (ETS2, APOE, PSEN1, DDC, and CD9) implicated mainly in the regulation of sarin-induced neuropathogenesis were further confirmed by Western blot and double-immunofluorescence assays. The regulome pathway suggests a new feasible mechanism by which sarin increases ETS2 expression and takes control over other genes involved in the neurodegenerative pathway. The overall data delineate an in vitro experimental model suitable for studying the neuropathology of cells and may provide novel insights into therapeutic interventions.
authors.affiliation: Departments of Anatomy and Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597.
authors.forename: Arjunan
authors.initials: A
authors.lastname: Pachiappan
authors.forename: Maung Maung
authors.initials: MM
authors.lastname: Thwin
authors.forename: Loke
authors.initials: L
authors.lastname: Weng Keong
authors.forename: Fook Kay
authors.initials: FK
authors.lastname: Lee
authors.forename: Jayapal
authors.initials: J
authors.lastname: Manikandan
authors.forename: Viswanathan
authors.initials: V
authors.lastname: Sivakumar
authors.forename: Ponnampalam
authors.initials: P
authors.lastname: Gopalakrishnakone
mesh_terms: D000110:Acetylcholinesterase / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002800:Cholinesterase Inhibitors / Q000494:pharmacology / Q000633:toxicity*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D046228:Microarray Analysis; D009474:Neurons / Q000187:drug effects*; D050787:Proto-Oncogene Protein c-ets-2 / Q000235:genetics / Q000378:metabolism*; D012524:Sarin / Q000494:pharmacology / Q000633:toxicity*; D015398:Signal Transduction / Q000187:drug effects*; D013997:Time Factors
chemical_list: D002800:Cholinesterase Inhibitors; C494069:ETS2 protein, human; D050787:Proto-Oncogene Protein c-ets-2; D012524:Sarin; D000110:Acetylcholinesterase