Study Of Toxic Effects And Mechanisms Of Baicalin Metal Complexes On SMMC-7721 Cells

Malignancies seriously affect human health,the research of anticancer drugs attracted widespread attention.Recent studies show that baicalin(BC)have some anti-tumor activity.However,little attention is paid to anti-tumor activity of baicalin metal complexe(BMC),and the mechanism of action is unclear.Therefore,in order to elucidating antitumor mechanism of BMC.The chemical,cell biology,molecular biology,electrochemical means multidisciplinary method has been used to synthesis BMC(Ni2+,Co2+,Cue+).We focus our study on the interaction of BMC on SMMC-7721 cell,and its DNA to explore the BMC and the strength of hepatocellular DNA binding capacity associated with its anti-tumor,which the main work is as follows:(1)The chelating ligand method has been used to synthesis BMC.The composition and structure of BMC were characterized by the elemental analysis,ultraviolet spectrum(UV),infrared spectrum(IR),mass and thermal gravitational(TGA)analysis.The results showed that three new kinds o·f BMC were successfully prepared,the molecular formulas of complexes are:Na2Ni(C21H16O11)2·10H2O,Na2Co(C2,H16011)2·8H20,Na2Cu(C21H16O11)2-8H20.(2)MTT,PI staining method and Annexin-V/FITC double staining method were used to analyze the effects of BMC on SMMC-7721 cell,such as cell proliferation,cycle and apoptosis,combined with morphological explore its anti-tumor activity.Fluorescence quantitative and flow cytometry were used to analyze the expression of BMC on Bcl-2,Bax mRNA and protein.The antineoplastic mechanism of BMC was explored comprehensively.The results showed that BMC restrained cell proliferation and cell apoptosis by stoping cell cycle,reduced the expression of Bcl-2 and increased that of Bax.The anti-tumor activity was:baicalin-copper(BC-Cu)>baicalin-cobalt(BC-Co)>baicalin-nickel(BC-Ni)>BC,showing the dose-response relationship.(3)Extracted hepatoma cells DNA as a target,cyclic voltammetry was utilized to study the interaction between BMC and DNA,to explore the interaction mechanism of both.The results showed that BMC and hepatoma cell DNA form a non-electroactive supramolecular compounds through electrostatic interaction and insert effect mixed-mode,binding number m=2,binding constant ?BC=2.77×106 L·mol-1,?BC-Ni=5.46x106 L·mol-1,?BC-co=7.74×106L·mol-1,?BC-cu= 1.21×107 L·mol-1.After BC complexes with metal ions its ability to bind to DNA significantly enhanced,the strength order of:BC-Cu>BC-Co>BC-Ni>BC.(4)We found the stronger binding capacity,the stronger anti-tumor activity by comparising the anti-tumor activity of BMC and the strength of the interaction between hepatocellular DNA.It indcated that the BMC-DNA complexes in cells exhibit ant-tumor activity by blocking DNA replications,inhibiting cell proliferation and leading cell apoptosis.This thesis successfully established associated model of BMC's anti-tumor activity and its with DNA binding ability,the relevant results provide new ideas for the design of new anticancer drugs.