Metabolism Analysis Of Doxorubicin In Breast Resistant Cancer Cell Line MCF-7/DOX

Classical pharmacokinetics(PK)can obtain the basic data of the body’s treatment of drugs,and study the absorption,distribution,metabolism and excretion(ADME)of drugs in the body.It is an important part of preclinical and clinical research of drugs.More and more new data show that in vivo pharmacokinetics cannot fully explain the pharmacological effects of drugs on target cells/organs.The internal treatment process of drugs and the concentration of the bound target are the decisive factors for observing the treatment response of target organs.Classical In this study,breast cancer cell line MCF-7 was selected as the target cell research material,and the method for extracting intracellular doxorubicin metabolites was optimized,and the UPLC-Q-TOF-MS/MS method was used to analyze the presence of doxorubicin in wild-type cells MCF-7 / WT and doxorubicin metabolites in MCF-7/DOX metabolites are different,and the structure of the different metabolites is deduced by multivariate multistage mass spectrometry Comparing the binding ability of the differential metabolites of doxorubicin in repeated cells with DNA through biocomputing simulation,and mining related enzymes involved in the formation of doxorubicin metabolites in combination with transcriptomics and proteomics data,in order to explain the anti-tumor drug doxorubicin The preventive mechanism of the element provides a reference.The main research work and results are as follows:(1)Study on the difference of doxorubicin metabolites in MCF-7/WT and MCF-7/DOX target cells.The wild strain MCF-7/WT of ductal ductal epithelial cells and the alternative strain MCF-7/DOX were selected as target cell research materials.Microbial sediment cultures containing doxorubicin were used to extract intracellular metabolic components and use UPLC to separate the extracts.Components were analyzed by high-resolution tandem mass spectrometry.There is a characteristic absorption at 480 nm and the relatively stable structure of the doxorubicin core structure m/z 321.By deriving its chemical structure through stepwise tandem mass spectrometry,two unreported trace doxorubicin metabolites M1(m/z606)and M2(m/z 641)were found.These two metabolites are completely different from the doxorubicin metabolites found in plasma,liver or kidney,suggesting that tumor-targeted cells have unique and diverse drug metabolism pathways for doxorubicin.(2)Computer simulation of molecular docking analysis.Comparing the binding ability of doxorubicin and metabolites M1 and M2 to DNA through molecular docking,the results show that compared with the binding site of doxorubicin and DNA,the binding sites of target cell metabolites of doxorubicin M1 and M2 and DNA all showed a certain degree of deviation,and their affinity with DNA decreased.It shows that doxorubicin metabolites in the target cell MCF7/DOX may be reduced by the anti-tumor pharmacological activity of the metabolites,which may be related to the anti-tumor drug resistance mechanism.(3)Prediction of metabolic enzymes related to doxorubicin metabolites.Based on transcriptome simulation analysis of the difference in gene expression between MCF-7 / WT and MCF-7 / DOX cells,401 signal-to-noise ratios(signal-to-noise ratio,SNR)> 4.0 were screened on the binding protein chip that can be compatible with doxorubicin Specific binding proteins,analysis of 100 targets related to doxorubicin metabolism,15 of which can be expressed as enzymes,including oxidoreductases,transferases,invertases,lyases and synthetases,including oxidoreductases and transferases.Some of these enzymes have been shown to be related to tumor formation or tumor drug resistance,which proves that it is feasible to screen drug resistance targets through transcriptomics combined with proteomics data.These targets can provide references for further research on the resistance mechanism of target cells to doxorubicin treatment.