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Induction of autophagy in human neuroblastoma SH-SY5Y cells by tri-ortho-cresyl phosphate

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Abstract

Tri-ortho-cresyl phosphate (TOCP) is an organophosphorus ester and has been widely used in industry. It is found that TOCP induced delayed neurotoxicity in humans and sensitive animal species. However, the mechanism of TOCP-induced neural cytotoxicity remains unclear. In this study, we studied whether autophagy is involved in TOCP-induced neural cytotoxicity in human neuroblastoma SH-SY5Y cells. We found that 0.5 and 1.0 mM TOCP treatment significantly increased the ectopic accumulation of microtubule-associated protein 1 light chain 3 (LC3)-immunopositive puncta, Beclin 1, and LC3-II/LC3-I levels in SH-SY5Y cells in a dose-dependent manner. Notably, by monodansylcadaverine staining method, we found abundant punctate fluorescent acidic vesicular organelles in TOCP-treated cells. Furthermore, ultrastructural observation under the transmission electron microscope indicated that the cytoplasm was occupied by autophagosomes in TOCP-treated SH-SY5Y cells. Thus, these results suggest that TOCP may induce autophagy, and autophagy may be involved in the development of TOCP-induced neural cytotoxicity.

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Abbreviations

AVOs:

Acidic vesicular organelles

ATG:

Autophagy gene

ECL:

Enhanced chemiluminescence

GFP:

Green fluorescent protein

LC3:

Microtubule-associated protein 1 light chain 3

MDC:

Monodansylcadaverine

PBS:

Phosphate-buffered saline

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

ST:

Starvation

TBS:

Tris-buffered saline

TOCP:

Tri-ortho-cresyl phosphate

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Acknowledgments

This work was supported in part by the grants from the National Natural Science Foundation of China (Nos. 81172712, 31071919), the Natural Science Foundation of Hunan Province, China (No. 11JJ6078), and the Youth Foundation of Education Bureau of Hunan Province, China (No. 09B087). We thank Mrs. X.-Q. Liu for her technical assistance with confocal microscopy.

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Laboratory of Molecular Toxicology, State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, 1-5 Beichenxi Road, Beijing, 100101, People’s Republic of China

    Ding-Xin Long, Pan Wang & Yi-Jun Wu

  2. School of Public Health, University of South China, Hengyang, 421001, People’s Republic of China

    Ding-Xin Long & Dan Hu

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  1. Ding-Xin Long
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  2. Dan Hu
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Correspondence to Yi-Jun Wu.

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Long, DX., Hu, D., Wang, P. et al. Induction of autophagy in human neuroblastoma SH-SY5Y cells by tri-ortho-cresyl phosphate. Mol Cell Biochem 396, 33–40 (2014). https://doi.org/10.1007/s11010-014-2139-7

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  • DOI: https://doi.org/10.1007/s11010-014-2139-7

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