[Investigation of trichloroethylene-induced effects on subcellular proteomes in L-02 hepatic cells].
OBJECTIVE
To put the insight into the trichloroethylene (TCE)-induced effect on the differential expression of subcellular proteins in human normal liver cell line (L-02).


METHODS
The membrane proteins and nuclear proteins of TCE-treated (8.0 mmol/L) group and controls were extracted by subcellular proteome extraction kit, respectively. The TCE-induced differentially expressions were analyzed by a two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization tandem time-of-flight spectrometry (MALDI-TOF-MS). Bioinformatics analysis was used to reveal the biological processes and predict transmembrane domains of differential expressed proteins. The expression of ATP synthase subunit beta (ATP5B), heterogeneous nuclear ribonucleoprotein H2 (hnRNP H2) and far up steam element-binding protein 1 (FUBP1) were measured under TCE treatment by Western blot.


RESULTS
After TCE treatment for 24 h in L-02 cells, 14 membrane proteins and 18 nuclear proteins were identified as differential expression. After treated with TCE in concentrations of 0, 2.0, 4.0 and 8.0 mmol/L for 24 h, the relative levels of ATP5B expression were 1.00±0.03, 1.21±0.14, 1.25±0.12 and 1.48±0.17 (F = 8.51, P = 0.007), the relative levels of hnRNP H2 expression were 1.00±0.09, 1.22±0.15, 1.43±0.21, 1.53±0.17 (F = 6.57, P = 0.015), respectively; the relative levels of FUBP1 expression were 1.00±0.11, 0.91±0.07, 0.73±0.04 and 0.67±0.03 (F = 15.81, P = 0.001), respectively, which were consistent with the results in proteomics. The bioinformatics analysis showed that the most dominant biological process were involved in RNA processing (10 proteins, P = 2.46×10(-6)), especially in RNA splicing (9 proteins, P = 1.77×10(-7)).


CONCLUSIONS
The exposure of TCE could alter the expression of membrane proteins and nuclear proteins in L-02 cells. These abnormal expressed proteins involved in RNA splicing would provide novel clues for further understanding of TCE-induced hepatotoxicity.
Jinan University College of Pharmacy, Guangzhou 510632, China.
Aibo
A
Huang
Email: huax_mail@126.com.
Hua
H
Xu
Wenxu
W
Hong
Zhixiong
Z
Zhuang
Email: bio-research@hotmail.com.
Jianjun
J
Liu
D015153:Blotting, Western; D002460:Cell Line; D004265:DNA Helicases; D004268:DNA-Binding Proteins; D022781:Hepatocytes; D034541:Heterogeneous-Nuclear Ribonucleoprotein Group F-H; D006801:Humans; D025261:Mitochondrial Proton-Translocating ATPases; D020543:Proteome; D040901:Proteomics; D012323:RNA Processing, Post-Transcriptional; D016601:RNA-Binding Proteins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014241:Trichloroethylene
C506526:ATP5F1B protein, human; D004268:DNA-Binding Proteins; C492258:FUBP1 protein, human; C116747:HNRNPH2 protein, human; D034541:Heterogeneous-Nuclear Ribonucleoprotein Group F-H; D020543:Proteome; D016601:RNA-Binding Proteins; D014241:Trichloroethylene; D025261:Mitochondrial Proton-Translocating ATPases; D004265:DNA Helicases