[Differential gene expressions in cells with low-dose hydrogen peroxide-induced adaptive response: a study with FDDRT-PCR].
OBJECTIVE
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).


METHODS
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.


RESULTS
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).


CONCLUSIONS
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.
Wei
Qin-zhi
QZ
Department of Toxicology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China. cnwei99@163.com
Zhuang
Zhi-xiong
ZX
D000222:Adaptation, Physiological / Q000235:genetics*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D042967:Electron Transport Complex I; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D009245:NADH Dehydrogenase / Q000235:genetics; D016877:Oxidants / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D018076:DNA, Complementary; D005456:Fluorescent Dyes; D016877:Oxidants; D019253:Proto-Oncogene Proteins c-bcl-2; D006861:Hydrogen Peroxide; D009245:NADH Dehydrogenase; D042967:Electron Transport Complex I; C427643:NDUFS4 protein, human