Trichloroethylene-induced downregulation of miR-199b-5p contributes to SET-mediated apoptosis in hepatocytes.

Trichloroethylene (TCE) is a ubiquitous toxicant widespread in our environment. Exposure to TCE can cause severe liver damage. In previous studies, we detected an abnormal elevation of SET (a protein encoded by the SETgene in humans) which was observed in human HL-7702 cells (L-02 hepatocytes) under the treatment of TCE. Moreover, further study indicated that SET acts as an important mediator in TCE-induced hepatocyte apoptosis. The major functions of SET have been elucidated, while the regulatory mechanism responsible for modulation of SET remains unclear. In this study, four major microRNA-related databases were used to screen and identify 6 candidate microRNAs involved in the regulation of SET. Subsequent verification indicated that miR-21 and miR-199b-5p were decreased in TCE-treated L-02 cells, suggesting that miR-21 and miR-199b-5p (miR199b for short) miR199b potentially regulate SET expression. Additionally, the dual-luciferase system revealed that only miR199b could directly bind to untranslated region (3'-UTR) of the SETgene. Modulation of SET by miR199b was verified through overexpression and knockdown of miR199b in L-02 cells. Assessment of apoptosis indicated that elevated miR199b attenuated TCE-induced apoptosis, while reduced miR199b enhanced it. In summary, this study suggests that in cultured hepatocytes, TCE-induced suppression of miR199b drives SET induction, which further mediates the response to TCE.

SET promotes H2Ak9 acetylation by suppressing HDAC1 in trichloroethylene-induced hepatic cytotoxicity.

Trichloroethylene (TCE) was widely used as an industrial solvent which could cause severe liver damage. The histone chaperone SET have been identified as an important mediator of TCE-induced hepatic cytotoxicity in our previous study; however, the underlying regulatory mechanisms remain poorly understood. In this study, we found a total of 136 histone acetylation sites involved in TCE-induced hepatic cytotoxicity with the technique of Triton-acid-urea polyacrylamide gel electrophoresis (TAU-PAGE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Importantly, 17 histone acetylation sites were revealed to be mediated by SET in TCE-induced cytotoxicity. The acetylation of histone H2AK9 (H2AK9ac) was further validated by Western-blot analysis. The data showed that TCE treatment increased the acetylation of H2AK9 in hepatic L-02 cell and decreased the one in SET-knockdown L-02 cells. Besides, levels of the histone deacetylases (HDACs, including HDAC1, HDAC2, and HDAC3) was also analyzed. Interestingly, the level of HDAC1 was aberrantly suppressed in TCE-treated L-02 cells while enhanced in SET-knockdown L-02 cells. To further explore the potential role of HDAC1 in SET-mediated hepatic cytotoxicity of TCE, we employed RNA interference (RNAi) to knockdown HDAC1 in both wide type L-02 and SET-knockdown cells. The results showed that the siRNA inhibition of HDAC1 increased the acetylation of H2AK9. Taken together, our data suggested that SET promoted the acetylation of H2AK9 via suppressing the level of HDAC1, which was involved in SET-mediated hepatic cytotoxicity of TCE.