| Objective: The aim of this study was to explore the antitumor effect of H+-K+-ATP inhibitor(proton pump inhibitor,PPI)and investigate the effect of OMZ on gastric cancer cell metabolism.Reveal the mechanism of OMZ in inhibiting gastric cancer.Methods: Cell counting kit-8(CCK-8)and cell clone formation assay were used to detect PPI inhibiting gastric cancer cell proliferation;cell wound healing assay and transwell invasion and migration assay were performed to detect the invasion and migration of gastric cancer cells;In MGC-803 and SGC-7901 gastric cancer cell lines,apoptosis related proteins such as Bcl-2(anti apoptotic protein),Bax(pro-apoptotic protein)and epithelial mesenchymal transition(EMT)related proteins such as Vimentin,E-cadherin and β-catenin were detected by Western blot.The antitumor effect of omeprazole was evaluated in vivo by a xenograft model of nude mice.PI staining were performed to detected the ability of PPI arresting cell cycle in gastric cancer cells and Annexin V-FITC/PI staining were performed to detect OMZ inducing apoptosis.Ca2+fluorescent probe(Fluo-3-AM)was used to determine cell calcium influx in SGC-7901 and MGC-803 gastric cancer lines.Metabonomics can detect intracellular small molecules of relative molecular weight less than 1000,and we used 1H-NMR-based metabonomics method for the quantitative analysis of intracellular metabolites.Metabolic analysis by extracellular metabolite in cancer cells help to research the anti-tumor effect of PPI and explore microenvironment change in SGC-7901 and MGC-803 gastric cancer lines,and seek the effect of PPI on metabolism pathway of gastric cancer.Results: CCK-8 assay and cell clone formation assay indicated that PPI can significantly inhibited gastric cancer cell proliferation.Cell wound healing assay and transwell invasion and migration assay indicated that PPI can significantly inhibited cell migration and invasion.The nude mice experiment in vitro proved that PPI had inhibited effect on gastric cancer cells,and the result had statistical difference(P <0.05).Western-blot showed that PPI can down-regulate of anti apoptotic protein Bcl-2 and up-regulate of proapoptotic protein Bax in SGC-7901 and MGC-803 gastric cancer cells were;the experiment also found that PPI can influence the expression of EMT related proteins,namely the upregulation of E-cadherin,downregulation of Vimentin and β-catenin protein expression.Metabonomic analysis showed that in MGC-803 gastric cancer cell lines,compared to the control group,valine,leucine,isoleucine,glutamine,glutamine,pyruvate and glucose use was significantly reduced in PPI group.Intracellular metabolites like Ethanol and Lactate accumulated in large quantities;metabolites such as myo-Inositol,Tyrosine and Phenylalanine were reduced in gastric cancer cell lines.Conclusion: Our results suggest that PPI inhibits the proliferation,the migration and the invasion of gastric cancer cells.PPI induces apoptosis of gastric cancer cells by increasing calcium influx,up-regulating Bax,down-regulating Bcl-2 protein and blocking cell cycle progression.PPI inhibits the invasion and migration of gastric cancer cells by inhibiting the Wnt/beta-catenin signaling pathway.At the same time,PPI inhibits the growth of gastric cancer cell by affect the utilization of amino acid and glucose of gastric cancer cells.The anti-tumor effect of PPI is the result caused by multiple factors. |
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