The Suppressed Effect And The Research Of Mechanism On The Tumor Cell Lines Of The Combination Of Cinobufotoxin And SB

Tumor is a kind of disease that multiple factors,multiple steps and mulitiple genes changed,so each medicine can play a part in only one or several target,if we can combine multiple inhibitors of tumor that will get better effect,and drug combination will become a tendency of medical science oncology.The ASCO meeting holded in US drew off a new viewpoint that target treatment,drug combination and multiple target medicine combination et al.Such method will bring better effect and profit. There are a great of versions about the mechanisms of tumor growth and carcinogenesis to date,and the gene phenotype modify and signal transduction have been study hot about the mechanisms of tumor growth and carcinogenesis.Cinobufotoxin is a effective cell differentiation antileptic,Sodium butyrate is histone deacetylase inhibitor.We conclude the possible mechanism of combining such two drugs after analyzing such two drugs mechanism of action:â‘ enhance the arrest of tumour cell cycle,arrest the tumor cell in G₁ stage;â‘¡inhibit the angiogenesis around the tumor tissue and metastasis;â‘¢regulate the expression of P21^WAF1,P53,CDK₄ gene and protein that related to the apoptosis and cycle;â‘£synergistic inhibit the signal transduction.We consider the essentiality and rationality to combining such two drugs according to the analysis and previous expriment experience.The reseach used the empirical methods:1,We researched the inhibitory rate of cell growth by MTT assay and detected the effect on the tumor cell lines of the combination of cinobufotoxin and Sodium butyrate.We sieved best concentration and action time.2,We detected the change of proteinum by Western-blot and detected the change of gene.3,We detected cell cycle and cell apoptosis by flow cytometry(FCM).We detected the mechanism of signal transduction of the combination of cinobufotoxin and Sodium butyrate.Results disclosed:1,We found the drug combination of Cinobufotoxin and SB can increase the apoptosis rate of SMMC-7721 and A549 cell,compared with Cinobufotoxin and SB only,the difference shows significant(P＜0.05),but the extent of Cinobufotoxin and SB under optimal concentration induced apoptosis isn't so obvious,drug combination increase only the apoptosis rate from 9-10%to 13-14%compared with Cinobufotoxin and SB only,it elucidate that the mechanims of the drug under optimal concentration is suppression of proliferation but not inducing apoptosis of tumor cells.2,Cinobufotoxin and SB can both arrest such two kinds of tumor cells SMMC-7721and A549 in G₁ stage to suppress the proliferation of cell.The drug combination of Cinobufotoxin and SB can arrest SMMC-7721 being77.13%and A549 being 78.32%cell cycle in G₁ stage obviously compared with Cinobufotoxin and SB only(P＜0.05).3,Cinobufotoxin and SB only can also enhance the expression of P53 and P21 protein,suppress the expression of CDK₄/CyclinD₁ For example of P21 protein in SMMC-7721,P21 protein is 0.5 when using SB alonly,after combination P21 protein is 0.65.The expression tendency of these protein is more obvious in drug combination group compared with SB and Gefitinib group(P＜0.05),the expression of P53,P21 gene and protein decreased,CDK₄/CyclinD₁ increased when the P38MAPK kinase activated by P79350,such expriment delinate the drug combination of SB and Gefitinib increase the expression of P53,P21,decrease the CDK₄/CyclinD₁ compound though P38MAPK signal transduction pathway.It certifies that P38MAPK signal transduction pathway affect te proliferation mediated by cell cycle,we can adjust the change of the cycle an apoptosis through changing the related gene and protein.The experiment in vitro proofed the suppressed effect on the tumor cell lines of the combination of cinobufotoxin and Sodium butyrate,and elucidated the mechanism of signal transduction of the combination of cinobufotoxin and sodium butyrate.To establish rationale for clinic.