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Research Article

Phosphoproteomic analyses of L-02 liver cells exposed to trichloroethylene

, , , , , , , , , & show all
Pages 459-466 | Received 18 Jan 2015, Accepted 31 Mar 2015, Published online: 27 May 2015
 

Abstract

Trichloroethylene (TCE) is an environmental and occupational toxicant that has been shown to cause serious hepatotoxicity. However, the mechanisms underlying the hepatotoxicity of TCE remain unclear. Previously, we identified several apoptosis-related proteins in TCE-induced hepatic cytotoxicity. This study is aimed to analyze the changes in phosphoproteins in L-02 liver cells exposed to TCE using iTRAQ labeling, IMAC enrichment and LC–MS/MS. We identified 1878 phosphorylation sites in 107 proteins and found that 20 sites in 16 phosphoproteins were differentially phosphorylated in L-02 cells after TCE treatment. Among these phosphoproteins, 20% were protein localization and formation processes-related proteins, 38% were metabolism-related proteins and 42% were cellular process-related proteins, including transcriptional regulation and biogenesis. Moreover, two phosphoproteins, 4E-BP1 (37T) and MCM2 (139S), were validated as TCE-induced alteration of phosphorylation at specific sites by Western-blot analysis. Taken together, our study demonstrated that TCE exposure changed the levels of multiple phosphoproteins in L-02 liver cells, and the functional analysis suggested that these differentially expressed phosphoproteins might be involved in TCE-induced hepatic cytotoxicity.

Declaration of interest

The authors do not have any potential conflict of interest or financial interests to disclose. This work was supported by the NSFC (National Natural Science Foundation of China) [81273126], the Key Project of Guangdong Natural Science Foundation [S2012020010903], the Project of Shenzhen Basic Research Plan [JCYJ20130329161137543, JCYJ20130329103949656, 201201027] and the Upgrade Scheme of Shenzhen Municipal Key Laboratory [ZDSYS20140509101335476].

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