Hyperoside Induces Apoptosis Of Gastric Cancer Cell Line MKN-45 And Its Mechanism

ObjectiveGastric cancer is the malignant tumor with high morbidity and mortality.At present,the treatment of gastric cancer is mainly surgical resection and postoperative chemotherapy,but the recurrence rate and metastasis rate are high,and there are many adverse reactions of chemotherapy drugs.Therefore,the research and development of anti-gastric cancer drug has become a hot research.Some studies have shown that hyperoside,this flavonoid compound could against lung cancer,pancreatic cancer,esophageal cancer and other malignant tumors.However,the mechanism of anti-gastric cancer MKN-45 cell line has not been reported.This article mainly discussed the effect of hyperoside on the proliferation and apoptosis of human gastric cancer MKN-45 cells,which could further investigate the antitumor effect of hyperoside.Methods1.Human gastric cancer MKN-45 cells were cultured in DMEM medium which contained 10% fetal bovine serum(FBS)in a incubator with saturated humidity at 37℃ and 5%CO2.2.The next day,after MKN-45 cells adherent,the experimental group was treated with Hyperoside 12.5,25,50,100,200μg/m L for 24,48 h,while the control group was not treated.3.After treated with Hyp,the cell activity was detected,the effect of Hyp on the proliferation of MKN-45 cells was observed and the suitable time and concentration of drugs were screened.4.Flow cytometry was used to detect the cell cycle changes and apoptosis rate of MKN-45 cells treated with Hyp for 48 h,which could observe the effect of hyperoside on apoptosis of MKN-45 cells.5.Western blot technique was used to analyze the expression of NF-κB P65,caspase-3 and Bcl-2 protein in MKN-45 cells treated with Hyp for 48 h,which could also observe the effect on hyperoside inducing apoptosis of MKN-45 cells.Results1.MTT results showed that MKN-45 cells were treated with different concentrations of Hyperoside(12.5,25,50,100,200μg/m L)for 24,48 h and treated with different concentrations of drugs at the same time.The lowest dose group(12.5μg/m L)compared with the control group,there was no significant difference(P>0.05).The proliferation rate of other drug treated groups decreased with the increase of drug concentration,and the difference was statistically significant compared with the control group(P<0.05).The effect of the same concentration of drugs in the experimental group was different at different time,and the difference was statistically significant in 50,100μg/m L group(P<0.05).The proliferation of MKN-45 cells was time-concentration dependent.2.Flow cytometry showed that the cell ratio of G0/G1 phase increased after Hyp treatment for 48 h.The cell ratio of S,G2/M phase decreased.Compared with the control group,the difference was statistically significant(P<0.01).MKN-45 cell cycle was blocked at G0 /G1 phase.3.Flow cytometry showed that the apoptosis rate of MKN-45 cells treated with different concentrations of Hyp for 48 h was significantly higher than that of control group(P<0.01).4.Western b Iot showed that the expression of caspase-3,Bax,IκBα increased and the expression of NF-κB p65,Bcl-2 protein decreased in MKN-45 cells treated with hyperoside for 48 h,and the difference was statistically significant(P<0.05),which further indicated that Hyp could induce apoptosis of MKN-45 cells.Conclusion1.Hyperoside inhibited the proliferation of MKN-45 cell line in a time and concentration dependent manner.2.The mechanism of hyperin inducing apoptosis in MKN-45 cell line might be related to blocking cell cycle,inhibiting NF-κB pathway,reducing the expression of NF-κB P65 and Bcl-2 protein and elevating the expression of caspase-3,IκBα and Bax protein.