加味柴芍六君方抑制IL-8介导的胃癌上皮间质转化的机制研究OA

Objective:To investigate the regulatory mechanism of modified Chaishao Liujun Prescription(柴芍六君方)on interleukin-8(IL-8)during epithelial-mesenchymal transition(EMT)in gastric cancer,and to provide new strat-egies for its treatment.Methods:The cytotoxicity of modified Chaishao Liujun Prescription on human gastric mucosal cells(GES-1)and its effects on the proliferation of gastric cancer cells(HCG-27)were detected by MTT assay.Scratch wound healing and Transwell invasion assays were used to evaluate its inhibitory effects on cell migration and invasion.An EMT model was induced by TGF-β1(10 μg/L),and an IL-8 overexpression model was constructed by lentiviral transfection.ELISA was used to measure the secretion levels of carcinoembryonic antigen(CEA),gastrin-17(G-17),and IL-8.The proteins and mRNA expression changes of EMT markers(E-cadherin,N-cadherin,Vimentin,Twist,Snail)and IL-8 were analyzed by Western blot and qRT-PCR.Immunofluorescence was used to observe the localization of IL-8 protein.Results:Compared with the blank control group,modified Chaishao Liujun Prescription(400 μg/mL)grop sig-nificantly inhibited the proliferation,migration,and invasion(P<0.01)of HCG-27 cells,the TGF-β1 group,the IL-8 group,and modified Chaishao Liujun Prescription(200,400 μg/mL)groups exhibited significantly elevated levels of CEA,G-17,and IL-8(P<0.01)of cells,and the IL-8 overexpression group exhibited significantly elevated levels of CEA,G-17,and IL-8(P<0.01)of cells.Compared with the IL-8 overexpression model control group,modified Chaishao Liujun Prescription(200,400 μg/mL)groups exhibited significantly reduced levels of CEA,G-17,and IL-8(P<0.05 or P<0.01)of cells,modified Chaishao Liujun Prescription(400 μg/mL)+overexpression group exhibited significantly downregulated the protein and mRNA expression of N-cadherin,Vimentin,Twist,Snail,and IL-8,while up-regulating E-cadherin(P<0.05 or P<0.01)of cells,and the TGF-β1 group showed upregulated the mRNA expression of Twist,while downregulating Il8,Ecadherin(P<0.05 or P<0.01)of cells.Compared with the TGF-β1 group,modi-fied Chaishao Liujun Prescription+TGF-β1 group showed downregulated the mRNA expression of Vimentin,Twist,and Snail,while upregulating E-cadherin and IL-8(P<0.05 or P<0.01)of cells.Immunofluorescence results indicated that,compared with the blank control group,the IL-8 overexpression group and the TGF-β1 group showed stronger fluores-cence intensity,whereas the modified Chaishao Liujun Prescription group attenuated IL-8 fluorescence signal of cells.Conclusion:By regulating IL-8 and its downstream signaling pathways,the modified Chaishao Liujun Prescription signif-icantly inhibited EMT in gastric cancer cells.