Effect On Erianin And 5-Fluorouracil On Cell Proliferation And Apoptosis In Human Gastric Cancer MGC-803 Cells

Objectives The effects of Erianin,5-fluorouracil and their comb-ination on the cell proliferation,apoptosis induction were observed by ta-king human gastric cancer MGC-803 cells as researching objects,And in-vestigated that if they induced apoptosis by regulate the expressions of Bax,Bcl-2,Caspase-3.The ultimate goal is to look for new gastric cancer drugs or promote the traditional 5-fluorouracil chemotherapy regimens.Methods(1)Gastric cancer cells were treated with Erianin or 5-Fu at different concentration(Erianin 0.977-250ng/ml,5-Fu 781.25-200000ng/ml)for different time(24h,48 h,72h),.Cells inhibition rates were detected with CCK-8 assay.(2) we used five groups to conduct the IC50 and combin-ation index according to the median-effect method,the groups were as follows:Erianin(0.977-250ng/ml),5-Fu(781.25-200000ng/ml),the combination(Erianin 0.977 ng/ml+5-Fu 781.250ng/ml,Erianin 3.906ng/ml+5-Fu 3125 ng/ml,Erianin 15.625ng/ml+5-Fu 12500ng/ml,Erianin 62.500ng/ml+5-Fu 50000ng/ml, Eria-nin 250ng/ml+5-Fu 200000ng/ml),a control group. all of the groups were treated with 48 h.(3)From the step2,we can gain the IC50 of the combination was 4329.044ng/ml,among that the concentration of Erianin was 5.404ng/ml and 5-Fu was 4323.640ng/ml.And these concentrations were used for the follow-up experiments.(4)The flow cytometer was used to analyze the effect of cell cycle.(5)The fluorescent microscope was used to observe the apoptosis of cells after treated with drugs.(6) The reverse transcription-polymerase chain reaction and Western blotting were used to analyze the expressions of Bcl-2,Bax and Caspase-3.Results(1)Both Erianin and 5-Fu could inhibit cell proliferation of human gastric cancer MGC-803 cells depending on time and concentration(p<0.05). we can get that the combination index was greater than 1 when the drugs combined in lower dose,but less than 1 when combined in higher dose.(2) The FCM told us that Erianin mainly arrest cell cycles in G2/M phase,while 5-Fu mainly arrest cell cycles in S phase. When compared to 5-Fu,the combination arrested less cells in S phase,but compared to Erianin,it arrested less cells in G2/M phase.(3) The fluorescent microscope was used to observe the apoptosis of cells.Through observing,we found that the apoptotic cells from the single drug group was much more than the control group,while the combination has the most apoptotic cells.(4)The RT-PCR proved that both the two drugs can induce the expressions of Bax,Bcl-2,and Caspase-3m RNA.The Western Blot proved that both the two drugs can promote the expressions of Bax,Caspase-3 and decrease the expression of Bcl-2,and the combi has stronger effect than the two single drugs(p<0.05).Conclusions:(1) In a certain concentration range,both Erianin and 5-Fu can inhibit cell proliferation of human gastric cancer MGC-803 cells depending on time and concentration.(2) The antagonism effect is only when the drugs combined in lower dose,When the combined dose increasing,the synergistic effect carry out.(3) The Erianin combined with 5-Fu may induce apoptosis through influencing the expressions of Bax,Caspase-3 and Bcl-2.