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[DNA damage and c-myc gene expression of NIH3T3 cell induced by superoxide anion]
1Department of Biophysics, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
Objective:
To study the DNA damage, membrane lipid peroxidation, and c-myc gene expression of NIH3T3 cell induced by superoxide anion(O2.-).
Methods:
The superoxide anion(O2.-) was produced by reactive system of xanthine-xanthine oxidase (X-XO) and the DNA-EB complex, malondialdehyde (MDA) content and expression of c-myc gene were measured by means of fluorescence, thiobarbituric acid (TBA) assay and DNA-RNA situ hybridization with digoxigenin labeled probes respectively.
Results:
The O2.- of high concentration(X: 20 micrograms, XO: 2 x 10(-3) U) reacted with isolated DNA from NIH3T3 cell directly which caused DNA damage obviously. When the O2.- of high concentration reacted with whole NIH3T3 cell, the amount of MDA increased and fluorescence intensity of DNA-EB complex decreased which mean lipid peroxidation of membrane and gene damage of the cell respectively. When the producing O2.- of high concentration reactive system existing FeSO4, the phenomena mentioned above showed obviously. Enough amount of catalase showed the inhibit effects, but it could not completely inhibit the DNA damage of whole cell. The c-myc gene expression was observed in the cell treated with O2.- of middle (X: 4 micrograms, XO: 4 x 10(-4) U) and high concentration, but it was not observed in the cell treated with O2.- of low concentration(X: 0.4 micrograms, XO: 4 x 10(-5) U).
Conclusions:
DNA damage of the whole cell induced by extracellular O2.- was able to act through the pathway of membrane lipid peroxidation in which hydrogen peroxide is important and the other pathways such as signal transduction of the cell might also be existed. The expression of c-myc gene of NIH3T3 cell induced by O2.- was related closely to the concentration of O2.-.