Paraoxon-Induced Protein Expression Changes to SH-SY5Y Cells
SH-SY5Y neuroblastoma cells were examined to determine changes in protein expression following exposure to the organophosphate paraoxon (O,O-diethyl-p-nitrophenoxy phosphate). Exposure of SH-SY5Y cells to paraoxon (20 μM) for 48 h showed no significant change in cell viability as established using a...
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Veröffentlicht in: | Chemical research in toxicology 2010-11, Vol.23 (11), p.1656-1662 |
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Sprache: | eng |
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Zusammenfassung: | SH-SY5Y neuroblastoma cells were examined to determine changes in protein expression following exposure to the organophosphate paraoxon (O,O-diethyl-p-nitrophenoxy phosphate). Exposure of SH-SY5Y cells to paraoxon (20 μM) for 48 h showed no significant change in cell viability as established using an MTT assay. Protein expression changes from the paraoxon-treated SH-SY5Y cells were determined using a comparative, subproteome approach by fractionation into cytosolic, membrane, nuclear, and cytoskeletal fractions. The fractionated proteins were separated by 2D-PAGE, identified by MALDI-TOF mass spectrometry, and expression changes determined by densitometry. Over 400 proteins were separated from the four fractions, and 16 proteins were identified with altered expression ≥1.3-fold including heat shock protein 90 (−1.3-fold), heterogeneous nuclear ribonucleoprotein C (+2.8-fold), and H+ transporting ATP synthase beta chain (−3.1-fold). Western blot analysis conducted on total protein isolates confirmed the expression changes in these three proteins. |
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ISSN: | 0893-228X 1520-5010 |
DOI: | 10.1021/tx100192f |