Establishment And Application Of Purine Nucleotide Metabolism Electrochemical Detection System

Objective To study the electrochemical behavior of the Hamster lung cells(V79cells),explore the effect of different factors on V79cell electrochemical signals, and establish purinenucleotide metabolism in cells.This system is used to research the influence that drugs inducedV79cell mutation on the hypoxanthine-guanine phosphoribosyltransferase (HGPRT) gene.Provide a theoretical basis for the establishment of HGPRT gene mutation detectioned byelectrochemical methods.Methods V79cells were used as experimental model, the electrochemical behavior of V79cells were studied by cyclic voltammetry, electrochemical signals of V79cells were tested byhigh performance liquid chromatography, gene locus mutation of V79cells were stydied byelectrochemical method and HGPRT gene mutation detection method.Results The pyrolysising liquid of V79cells have two electrochemical peaks that are theoxidation of guanine and xanthine (0.7V) and the oxidation of adenine and hypoxanthine (1.03V). The best conditions are that constant temperature of50℃for30min with pH7.4.Theresults show, the peak area of signal I in treat group obviously higher than that of control groupafter drug treating of the fourth day and the peak area of signal I on fifth day in theconcentration of1.6mg·mL-1on the fourth day. The study of different treatingconcentrations of the EMS found that the mutation rate increased following the increase ofcondition of EMS and the difference of peak area between dosing group and the controlgroup increased with the increase of doses between the concentration of0.7and theconcentration1.6mg·mL-1.Conclusion The two obvious signal peak have been tested in V79cells throughelectrochemistry method, the two signal peaks are used indicator. Establishing purineelectrochemical detection system within the cells and used in the stuidy of HGPRT genemutation. The results have showed that it’s consistent with the result of conventional HGPRTgene mutation. The results of electrochemical detection have shown that genen mutation hashigher electrochemical signals than control group after treating of the fourth day, which canshort the testing time of the conventional HGPRT gene mutation for up to15days and improvesthe objectivity of measuring results.