The Study Of Yangzheng Sanjie Decoction Regulating Gastric Cancer Growth And Signaling Pathway

ObjectiveGastric cancer is an upper gastrointestinal tumor disease with high morbidity and high mortality rate in China.Many studies have shown that Chinese medicine has unique advantages in the treatment of gastric cancer and has broad application prospects.The previous experiments in vitro of research group showed that traditional Chinese medicine compound Yangzheng Sanjie decoction(YZSJD)-containing serum could regulate the growth of gastric cancer cells through miR-7,inhibit its proliferation and induce its apoptosis.On this basis,this study further verified the effect of YZSJD extract powder on the proliferation and migration of gastric cancer cells,and explored the possible cell signaling pathways.Methods1.Human gastric cancer cell line MKN-45 were uesd to establish a subcutaneous tumor formation model in nude mice.All mice were randomly divided into three groups:model control group,YZSJD and positive control group.During administration,general condition of nude mice were observed.Meanwhile the volume and weight of tumors in each group were measured to verify the inhibitory effect of YZSJD on transplanted tumor.2.Cell proliferation was detected by CCK-8 method and plate clonal formation experiment,cell migration was detected by scratch healing experiment after treating human gastric cancer AGS and MKN-45 cell lines with YZSJD extract powder in vitro.3.After the intervention of YZSJD in vitro,the expression of protein EGFR,the expression and its phosphorylation of ERK and AKT protein in gastric cancer cells were detected by Western blotting.4.Western blotting was used to detect the expression of p65 and p50 proteins in gastric cancer cells after treatment with YZSJD,which explored the effect of YZSJD on NF-κB signaling pathway.Results1.After 28 days of administration,the tumor suppressor rate of nude mice were 59.57%and 48.94%,respectively,in the positive control group and YZSJD group.Compared with the model control group,the growth of the transplanted tumors of the two groups was significantly inhibited(P<0.01).2.After different concentrations of YZSJD were applied to gastric cancer AGS and MKN-45 cells for 24 h,48 h and 72 h,the survival rate of gastric cancer cells was significantly decreased,and cell proliferation was significantly inhibited(P<0.05).After AGS cells were treated with the drug concentration of YZSJD(0.2,0.4,0.6,0.8,1 g · L-1)for 48 h,the clone formation rates of the control group and YZSJD group were:51.04%,49.38%,38.92%,30.92%,19.33%,5.63%,respectively.After 48 hours of intervention in MKN-45 cells,the clone formation rates of the control group and YZSJD group(0.4,0.8,1.2,1.6 g · L-1)were:38.83%,32.23%,17.80%,6.33%,0.70%.When the drug concentration was 2 g·L-1,it is basically impossible to form a cell colony.Compared with the control group,the clonal formation rate of gastric cancer cells in YZSJD group decreased significantly(P<0.05),and decreased gradually with the increase of the concentration of administration.3.After treatment of gastric cancer cells with YZSJD for 24 h and 48 h,the migration ability of cells was significantly lower than that of the control group(P<0.05).4.The drug concentration of YZSJD(1,2,4 g · L-1)was used to intervene in AGS cells,and 2,4,8 g · L-1 of YZSJD was used to intervene MKN-45 cells,the expression of EGFR protein in gastric cancer cells was significantly decreased(P<0.05).Compared with the control group,the phosphorylation level and expression of ERK protein in MKN-45 cells were down-regulated(P<0.05).After treatment of AGS cells with 2 g · L-1 of YZSJD for 6 h,12 h,and 24 h,the phosphorylation level of ERK protein was significantly down-regulated(P<0.05).Treated with 2 g · L-1 of YZSJD in AGS cells and 4 g · L-1 of YZSJD in MKN-45 cells respectively for 6 h,12 h,24 h,48 h,the phosphorylation level of AKT protein in gastric cancer cells was significantly down-regulated from the beginning of 24 h(P<0.05).5.After treatment with YZSJD for 48 h,the expression of p65 protein in AGS and MKN-45 cells were significantly decreased(P<0.05),and there was no statistically significant difference in the expression of p50 protein.ConclusionYZSJD significantly inhibits the growth of gastric cancer xenografts in nude mice and can reduce the proliferation and migration of gastric cancer cells in vitro intervention.YZSJD may exert anti-cancer effect by inhibiting the expression of EGFR protein and its downstream ERK and AKT signaling pathways,blocking the NF-κB signaling pathway in gastric cancer cells.