| With the wide application of chemotherapy,the tolerance of cancer cells to drugs is increasing,so that the combined application of drugs has become an inevitable trend.Polyphenols have a good therapeutic effect in the treatment of cancer and have small side effects.Anthracycline anticancer drugs have a wide range of clinical applications and are one of the commonly used chemotherapy drugs,but severe cardiotoxicity limits their application.By studying the interaction between polyphenol-anthracycline anticancer drug synergy system and human serum albumin at different temperatures,co-solute and pH and in vitro cytotoxicity determination,the mechanism of action of drug and human serum albumin and the relationship between the two drugs simultaneously interacting with HSA was determined.The experimental results can provide a theoretical basis for the combined application of clinical drugs.On the basis of sufficient literature research,combined with the actual situation of the laboratory,a variety of spectroscopy methods combined with in vitro cell experiments were used to study the interaction of polyphenol-anthracycline anticancer drug synergy system with HSA at different temperatures,co-solute and pH.This work is funded by the National Natural Science Foundation of China(No.21473085)and is divided into the following four parts:Firstly,the interaction of epigallocatechin gallate(EGCG)and anthracyclines with HSA at different temperatures and co-solutes was investigated by spectroscopy methods and cytotoxicity experiments.The thermodynamic parameters such as binding constant,number of binding sites,enthalpy change and entropy change were obtained.The mechanism of interaction between drug and protein,the interaction force and the protein structural change were determined.Moreover,the synergistic cytotoxicity of EGCG and DOX/EPI as well as their complexes with HSA were discussed.Secondly,spectroscopy methods and cytotoxicity experiments were conducted to study the interaction between oxidized resveratrol(OXY)and mitoxantrone(MIT)synergistic system and HSA,the effects of temperature and pH on binding were discussed.According to the experiment data,the binding sites of the two drugs and the configuration changes on HSA were obtained.Cytotoxicity to drug Hela cells was obtained by cell experiments.The results show that the binding of OXY and MIT to HSA is competitive,and the drug cytotoxicity is enhanced after forming a complex with HSA,and these two drugs produce a synergistic effect on Hela cells.Thirdly,spectroscopy methods were combined with in vitro cell experiments to study the interaction of piceatannol(PIC)and MIT with HSA at different temperatures and pH.The binding mechanism,the number of binding sites,the enthalpy change and the entropy change in the binding process were determined by fluorescence spectroscopy.The secondary structure and particle size of HSA were obtained by circular dichroism spectroscopy and dynamic light scattering.The effect of drugs on Hela cells was discussed by cell experiments.The results showed that the toxicity of Hela cells was enhanced after the formation of PIC and HSA complexes,PIC and MIT applied together were produced synergistic effects.Fourthly,the interaction between PIC/doxorubicin(DOX)and HSA was studied by spectroscopy methods and in vitro cell experiments.Experiment datas have shown that PIC and DOX bind to the same site and competitive binding of HSA.Comparing the changes of thermodynamic parameters between single drug and two drugs,it indicates that the presence of one drug weakens the binding of another drug to HSA.At the same time,the effects of two drugs on Hela cells were discussed.In addition,the thermodynamic parameters and cytotoxicity of polyphenols and anthracyclines combined with HSA were compared.It was found that the difference in the position and structure of the substituents resulted in different performances in terms of binding ability and cytotoxicity. |
PDF Full Text Request