Study On The Relationship Of The Structure-effect-toxicity Of Bufadienolides Inhibiting Tumor Cells Proliferation

The tumor was second only to cardiovascular disease caused humanity to dead,recent years,the incidence and mortality of China’s malignant tumor were on the rise.Existing antitumor drug treatment effect was not completely satisfying.So,looking for new anti-cancer drug candidates still was to have the important meaning.Bufadienolides,from Traditional Chinese medicine toad,had significant steroid anti-tumor effect.But such compounds in which the tumor’s most development value,was not clear.Especially the study of the relationship on the structure-effect-toxicity of bufadienolides inhibiting tumor cells proliferation was very little.This paper researched the most sensitive cell lines for bufadienolides,and the relationship between chemical structure and inhibiting tumor cell proliferation activity and toxicity of ten kinds of bufadienolides.The main contents included the following several parts:The literature reviewed the chemical composition,antitumor activity and molecular mechanism of the Chinese traditional medicine toads.The evaluation of the sensitivity to tumor cells of the bufadienolides.Selected cell lines SMMC-7721,SGC-7901,HT-29,Hela,Tca-8113,SK-OV-3,A431,MGC-803,A549;and rat heart muscle cells H9C2,liver cell L02,guinea pigs splenic cell for model experiment in vitro,MTT method to determine the inhibition of cell proliferation,to screen the most sensitive cell line to bufadienolides.The results of the sensitive cell line showed that:HT-29<A431<A549<MGC-803<SMMC-7721<SGC-7901<SK-OV-3<Tca-8113<Hela.Hela cell was the most sensitive cell line to bufadienolides,Tca-8113 cells was the second,and the sensitivity of normal cells was the worst.The evaluation of the activities inhibited tumor cell proliferation to ten bufadienolides.Selected cancer cells Hela,Tca-8113,and SK-OV-3,MTT method to determine antitumor activity of the bufadienolides.The antitumor activity of 10 bufadienolides to Hela cell line was that:Bul>Btl>Arg>Hel>Ctl>Cbl>Deb>Tel>Gtl>Rbg;the order for the antitumor activity of 10 bufadienolides to Tca-8113 cell line was that:Bul>Cbl>Btl>Arg>Tel>Deb>Hel>Ctl>Gtl>Rbg;the order for the antitumor activity of 10 bufadienolides to SK-OV-3 cell line was that;Bul>Cbl>Btl>Deb>Arg>Tel>Hel>Ctl>Gtl>Rbg.The overall results showed that the antitumor effect bufalin was the best,bufotalin and cinobufagin were the second,but the resibufogenin has no significant inhibiting tumor cell proliferation.The relationship between chemical structure and the inhibiting tumor cell proliferation activity of bufadienolides.10 bufadienolides were divided into two kind of mother nuclear structure,A-type and B-type,compared the inhibiting tumor cell proliferation activity;And then in the same nuclear structure,chemical structure were divided into various kinds of types,compared the tumor cell proliferation inhibitory activity again.In Hela cell line,it indicated that the chemical structure of A-type nucleus with 14-epoxy was significantly lower than the structure of B-type nucleus which withl 4-hydroxy.In the structure of A-type nuclear structure,the compounds with more hydroxy or acetyl group of bufadienolides can enhance their cytotoxicity;But in the B-type nuclear structure,It was just the opposite.At the same time,took Tca-8113 cell for in vitro model,this paper investigated and found that the conclusion was the same.The biological affinity of bufadienolides on the tumor cell.Cell extraction-UPLC-QTOF determination the biological affinity of ten bufadienolides on Hela cells,and found there were no direct correlation between the biological afinity and its antitumor vigor,concluded that the drug biological compatibility,in addition to the specific affinity and nonspecific affinity.Analyzed the correlation between the bioafinity rates and the properties computed from structure of bufadienolides,to study the effect of affinity nonspecific factors.Foundhe coefficient of correlation between XLogP3,H-Bond Donor and Topological Polar Surface Area with the decrease of bufadienolides in cell supernatant after affinity,lipid compounds,it was conclused that bufadienolides had different levels of affinity with Hela cells,which correlate with their Properties Computed from Structure.Compounds’ nonspecific attribute indirect impact on biological activity.The relationship between chemical structure and toxicity of bufadienolides.Select Hela cell line and SK-OV-3 cell line for vitro model,with sodium ions fluorescent probes S6901for indicator,investigated the effect of bufadienolides to free sodium levels in cells,explored the toxicity of bufadienolides to tumor cells,thus,screening the good effect and safety Compounds.Bufadienolides could induce the increasing of free sodium inside cells,it has positive correlation;There was no relevance between the activities inhibiting tumor cell proliferation and free sodium ions inside the cell,similarly,it figured out that there was no direct correlation between the activities inhibiting tumor cell proliferation of bufadienolides and its drug toxicity,thus,search out better and safe antitumor drugs preliminary.The preliminary exploration of protein electrophoresis.Selected Hela cells in vitro studies,and added bufadienolides into cultured Hela cells with different concentration,extracted cell plasma membranes protein for the SDS-PAGE electrophoresis,preliminary explored the influence of bufadienolides to tumor membrane protein.The results found that there’s obvious difference during dosing group and blank group,different dosing concentrations,too.It intuitively showed that bufadienolides influent the cytoplasm membrane protein of tumor cells,to lay the foundation for the research of proteomics in inhibiting tumor of bufadienolides.The preliminary exploration of endogenous metabolites inside cells effected by bufadienolides.Selected SK-OV-3 cells in vitro studies,and setted model control group and given medicine group,detected by UPLC-QTOF,and principal component analysis and orthogonal partial least variance discriminant analysis.The results found that there’s obvious difference during the dosing group and blank control group.Experiments could be the preliminary conclusion,bufadienolides could lead to the change of endogenous metabolites inside SK-OV-3 cells.It intuitively lay the foundation for the research of metabolites in inhibiting tumor of bufadienolides.