Study On The Cell Electrochemical Mechanism And The Method Of Measuring The Environmental Estrogens Based On The Voltammetric Behavior Of The MCF-7 Cell In Vitro

This paper studies the voltammetric response of the MCF-7 cells and the PC-3 cells as the model cell by using electrochemical method, HPLC method and chemometric analysed the kinds and the concentration of the electrochemical active specises of the cells, and explained the mechanism on the origin of the voltammetric response; By comparison of the electrochemical behaviors of the cell secretion and the cytoplasm, the electrochemical method of measuring active of the environmental estrogens based on the electrochemical behaviors of the cytoplasm was built. The electrochemical method are rapid, highly sensitive, and inexpensive. The results are as follows:1. The mechanism on the origin of the voltammetric response of the cells. The electrochemical behaviors of the living cell suspension and the cytoplasm were studied, and the results were found that the electrochemical response of the living cell suspension was given by the oxidation of guanine and xanthine in the cell secretion, not by the cells. Ultrasonication and heat treatment were carried out for obtaining the cytoplasm, whose voltammeric response was stronger than that of the cell suspension. The voltammeric response of the cytoplasm obtained by heat treatment was stronger than that by ultrasonication due to that the concentrations of guanine and xanthine in the cytoplasm obtained heat treatment were higher than that by ultrasonication.2. Building the system of evaluating the active of the cells by the electrochemical method. To suggest the relation between the electrochemical signal and the cell viability, the effect of cytotoxicity of taxol, luteolin, quercetin, isoquercitin and rutin on the cell viability was studied by electrochemical method. The result obtained by using the electrochemical method displayed cytotoxic tendencies similar to those by the conventional in vitro drug MTT and flow cytometry sensitivity tests, suggesting that the method of recording voltammetric measurements described above was credible. The altered response was driven by the decreased concentration of xanthine and guanine in the cytoplasm. The relation between the voltammeric behavior of cells and intracellular catabolism was interprected.3. Establishment the method of measuring environmental estrogens based on the cell electrochemistry. Comparative studies of electroactive components and voltammetric behaviors different between the MCF-7 cell without estrogen and the MCF-7 cell with estrogen showed that the concentration of the guanine and xanthine bases in the cytoplasm decreased, which resulted in the decrease of secretion ability of the MCF-7 cell without estrogen decreased, and these result in the decreasing of the voltammetric response of the cells, impling that the cell vaibility decreases. The effect of estradio. nonylphenol.bisphenol on the proliferation of the MCF-7 cells without estrogen.The results showed that dosage-dependen and time-dependen was present and the order of the estrogen effect is as follow: estradio>nonylphenol>bisphenol; The effect of environmental estrogens on the proliferation of the MCF-7 cells was studied by the electrochemica method, and the result above was similar to those by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method, suggesting that voltammetric measurements was credible.