Differential gene expressions in cells with low-dose hydrogen peroxide-induced adaptive response: a study with FDDRT-PCR / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify differentially expressed genes in human embryo lung fibroblasts MRC-5 with adaptive response induced by low concentration of hydrogen peroxide (H(2)O(2)) using fluorescent differential display-RT-PCR (FDDRT-PCR).</p><p><b>METHODS</b>The dose-effect pattern of H2O2 toxicity was determined using MTT assay, and the dose of 0.088, 0.88, 8.8, 88 micro;mol/L was defined as the low concentration, and 1100 micromol/L as the high concentration. Adaptive response model was established in MRC-5 cells verified using LDH release and cell apoptosis analyses. Differentially expressed genes in the cells with exposure to different doses of H(2)O(2) were detected by FDDRT-PCR, and some of the differentially displayed genes were determined using real-time quantitative PCR.</p><p><b>RESULTS</b>Cells challenged with high-concentration H(2)O(2) for 1 h after H(2)O(2) pretreatment at low concentrations for 24 h resulted in lessened toxic effect in comparison with direct high-concentration H(2)O(2) exposure. The adaptive response of the cells was most obvious with H(2)O(2) pretreatment at 0.88 micromol/L. Altogether 60 differentially expressed genes were detected with FDDRT-PCR in different treatment groups, and 5 of them were identified and verified, including 1 unknown gene and 4 known genes (bcl-2, EIF3S5, NDUFS4 and RPS10).</p><p><b>CONCLUSION</b>According to the results of FDDRT-PCR, the genes bcl-2, EIF3S5, NDUFS4 and RPS10 can be involved in H(2)O(2)-induced adaptive response of the MRC-5 cells.</p>

Differentially express genes in human embryonic lung fibroblasts with damage tolerance induced by low-dose hydroquinone / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the differentially expressed genes in human embryonic lung fibroblasts (HELF) induced by small-dose hydroquinone (HQ) using fluorescence differential display-PCR (DD-PCR).</p><p><b>METHODS</b>According to the dose-effect relation of HQ toxicity we established previously, HQ dose that did not induce observed cell damage or proliferation arrest was defined as low dose (100 pmol/L), and that causing obvious cell damage as the high dose (100 micrommol/L). The cells were then treated with low or high dose of HQ, or exposed to high-dose HQ following pretreatment with low-dose HQ for some time, respectively. Fluorescence DD-PCR was performed and 33 differentially expressed genes were identified in the cells with different treatments, and 8 of the identified genes were amplified, cloned, sequenced and blasted.</p><p><b>RESULTS</b>Seven of the 8 amplified genes were unknown genes, and the left one was identified as a known gene highly homologous to that encoding Homo sapiens Rap1 interacting factor 1 (RIF1).</p><p><b>CONCLUSION</b>Low-dose HQ can induce damage tolerance in HELF, and identification of the differentially expressed genes may provide valuable sight into the mechanism of HQ-induced damage tolerance.</p>

Study on differential proteomic expression in human liver cells stimulated by trichloroethylene with proteomics / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the differential proteomic expression in human liver cells L-02 induced by different dosages of trichloroethylene (TCE).</p><p><b>METHODS</b>Human liver cells L-02 were treated with different concentrations of TCE and the solvent control (dimethylsulfoxide). The total cellular proteins were separated using 2DE and visualized with silver staining after TCE treatment. The images were analyzed with Image Master 2D Platinum 5.0 analysis software. The differentially expressed protein spots were identified by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-TOF-MS).</p><p><b>RESULTS</b>Fifteen protein spots with significant difference were found, and went upward or downward or disappeared after the stimulation of TCE with different dosages, which indicated that TCE induced the change of the proteomic expression in the liver cells. The mass spectrum identification and the IPI human database retrieval were used for identifying 9 proteins related to the L-02 Liver cells induced by TCE.</p><p><b>CONCLUSION</b>The result provides an insight to TCE-related molecular mechanism and which might be useful for further study of the TCE-associated proteins and molecular markers.</p>

Two-dimensional electrophoresis and mass spectrometry analysis on effects of trichloroethylene on protein of L-02 liver cells / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the effects of trichloroethylene (TCE) on the protein in L-02 cells in vitro.</p><p><b>METHODS</b>Thiazolyl blue and Trypan blue tests were used to investigate the cytotoxicity of TCE to L-02 liver cell. The 2-D electrophoresis was used to analyse the expression of proteins in L-02 liver cells. The differentially expressed protein spots were identified by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-TOF-MS).</p><p><b>RESULTS</b>When the concentration of TCE exceeded 30 micromol/L, there was distinct cytotoxicity to L-02 cell (P < 0.05). Selected 40 micromol/L to treat L-02 liver cells and analyze the differential proteome expression, the results showed that the expression level of 37 protein spots was up-regulated and 15 protein spots was down-regulated. And 15 proteins were identified by MALDI-TOF-TOF-MS.</p><p><b>CONCLUSION</b>TCE can change the proteome expression of L-02 liver cell. It should provide the fundamental information to identify proteins related to TCE in further study.</p>