Effects Of Tolfenamic Acid On Proliferation And Apoptosis Of The Human Gastric Cancer AGS Cell Line

BackgroundTolfenamic acid(2-[(3-chloro-2-methyl phenyl)amino]benzoic acid)is a new type of nonsteroidal anti-inflammatory drugs,the action mechanism is blocking the generation of prostaglandins from arachidonic acid via inhibition of cyclooxygenase-1 and antipyretic analgesic.At present,tolfenamic acid have been used in clinical diseases such as rheumatoid arthritis,migraine treatment.In recent years,tolfenamic acid was found that it can inhibit tumor cell growth,induction of apoptosis,cell signal transduction,regulation proto-oncogenes and tumor suppressor gene activity and inhibition of tumor angiogenesis by domestic and foreign scholars from various aspects.Tolfenamic acid has antitumor effect of tolfenamic acid on pancreatic cancer,esophageal cancer,colorectal cancer and other digestive system malignant tumor,but the tolfenamic acid on the antitumor activity and function of gastric cancer has not been reported.So in this study,tolfenamic acid on the antitumor activity and function of the gastric cancer cells were detected by the MTT method of leading to cell proliferation,FITC Annexin V/PI double staining test cell apoptosis rate,Hochest 33258 AGS cell nuclei and Western Blotting method.ObjectiveTo investigate the effects of tolfenamic acid on proliferation and apoptosis of the human gastric cancer AGS cell line.Methods1.The effect of tolfenamic acid on AGS gastric cancer cell apoptosis rate were investigated by FITC Annexin V/PI staining and flow cytometry;2.The effect of different concentrations,different action time of tolfenamic acid on AGS gastric cancer cell growth was tested by MTT assay;3.The effect of tolfenamic acid on AGS gastric cancer cell apoptosis were investigated by Hochest 33258 nuclei through tolfenamic acid treatment of AGS cells,and observe the difference apoptosis by the fluorescence microscope;4.Western Blotting method were used to investigate the relative expression quantity of Bel-2,Bax protein and Bel-2/Bax ratioResults1.After 24 hour the apoptosis rate of the control group,50?mol/L?100?mol/L.200?mol/L tolfenamic acid group were 15.32±2.26.29.87±4.76?54.821±4.49.72.3±5.53).Compared with control group,tolfenamic acid of different concentration on cell apoptosis of treatment group was obviously increased(P<0.05),and the apoptosis rate increased as the drug concentration gradient increases(P<0.05).2.When AGS gastric cancer cells were treated by 50?mol/L?100?mol/L?200 ?mol/L tolfenamic acid group for 24h,48h,72h,The results of MTT assay indicated that compared with negative control group,cell proliferation ability of tolfenamic acid groups were significantly inhibited(P<0.05).on the other hand,as the increase of drug concentration,the inhibition ability were enhanced(P<0.05).3.The apoptotic cells in the control group number was2.32±0.54,and the apoptotic cells in the tolfenamic acid groups number was 9.17±1.38,(P<0.05).Compared with the control group,tolfenamic acid groups of cells were treated after 24 hours,Hoechst 33258 nuclear staining showed that AGS cells displayed significant apoptosis,with the more frequent appearance of condensed and fragmented nuclei.4.Western Blotting method were used to detect the relative expression quantity of Bel-2 and Bax protein of 50?mol/L?100?mol/L?200?mol/L tolfenamic acid groups,the result showed that Bcl-2 was 1.42±0.11?0.95±0.14?0.63±0.08,and Bax was 0.28±0.03?0.57±0.05?0.86±0.09,(P<0.05).As the increase of drug concentration,the Bcl-2/Bax protein ratio were decreased.Conclusion1.The antitumor effect of tolfenamic acid can be achieved by inhibiting growth of AGS gastric cancer cells proliferation and promoting cell apoptosis.2.The role of Tolfenamic acid promoting AGS gastric cancer cell apoptosis is related to down-regulation of the Bcl-2 protein expression and up-regulation of Bax protein expression.