| Research background and purposeGastric cancer ranks second in the incidence of malignant tumors in China and seriously threatens the health of the people.Due to the atypical symptoms that are difficult to detect early,most patients with gastric cancer are diagnosed at the advanced stage.Drug treatment is the main treatment of advanced gastric cancer,but there is no standard treatment plan,and the progress of targeted therapy has achieved little effect.It is urgent to discover new mechanisms and new targets for gastric cancer treatment.miRNAs participate in tumorigenesis by regulating mRNA,and have important regulatory effects on the proliferation,apoptosis,and drug sensitivity of malignant tumor cells,including gastric cancer.Earlier research by our group found that esomeprazole can inhibit the proliferation,invasion and migration of gastric cancer cells,induce their apoptosis and enhance the sensitivity of chemotherapy drugs.However,the mechanism by which esomeprazole regulates the biological behavior of gastric cancer cells is unclear.Therefore,this study explored the effect of esomeprazole on the expression profile of miRNA in human gastric cancer AGS cells,and performed functional prediction analysis of differentially expressed miRNAs through bioinformatics methods to explore the possible mechanism of action of esomeprazole on gastric cancer cells.The preliminary verification of the expression and role of differentially expressed miRNAs in human gastric cancer AGS cells and SGC-7901 cells has laid a foundation for further revealing the mechanism of esomeprazole's anti-gastric cancer cell effects by regulating miRNA.Research methodThe effect of esomeprazole on the inhibition of gastric cancer cell proliferation was detected by the CCK-8 method.The effect of esomeprazole on miRNA expression profile of human gastric cancer AGS cells was detected by gene sequencing method,and differentially expressed miRNAs were screened.Cluster analysis and functional prediction analysis of differentially expressed miRNAs were investigated by bioinformatics software.The expression of miRNA-22-3P and miRNA-22-5P in AGS cells and SGC-7901 cells under the action of esomeprazole was verified by qPCR.Transient cells transfected with small molecule inhibitors were used to construct transient miRNA-22-3P low-expressing gastric cancer cell lines,and its effect on the inhibition of esomeprazole proliferation was observed by the CCK-8 method.Statistical analysis was performed by SPSS 25.0 statistical software package.P<0.05 was considered with statistically significance.Research results1.CCK-8 test results showed that esomeprazole at different concentrations acted on human gastric cancer AGS cells for 48 hours,and the cell proliferation inhibition rate increased with the increase of esomeprazole concentration,the difference was statistically significant(P<0.05).2.G ene sequencing results showed that there were significant differences in miRNA expression profiles before and after esomeprazole.Among 7350905 miRNA transcripts,there were 199 differentially expressed miRNAs,of which 71 were up-regulated and 128 were down-regulated.Among them,hsa-miR-4758-3p and hsa-miR-548d-3p are sorted by the most significant difference in up-regulation and down-regulation of miRNAs;hsa-miR-22-3p and hsa-miR-3529-3p is the most significant miRNA with up-and down-regulation of differential expression,respectively.3.GO analysis was performed on differentially expressed miRNAs,and it was found that in biological process clustering,signal transduction,protein phosphorylation,and intracellular signal transduction were mainly involved.In cell component clustering,cytoplasm,nucleus and Plasma membrane were mainly involved.In molecular functional clustering,protein binding,metal ion binding and ATP binding were mainly involved.4.KEGG Pathway analysis was performed on the differentially expressed miRNAs.The results showed that the four pathways significantly related to tumors were:cAMP signaling pathway,MAPK signaling pathway,pathway in cancer,and cGMP-PKG signaling pathway.5.Quantitative PCR verification results showed that miRNA-22-3P and miRNA-22-5P were significantly up-regulated in human gastric cancer AGS cells and SGC-7901 cells treated with esomeprazole(P<0.05).The difference was more pronounced in-7901 cells.6.CCK-8 test results show that compared with the control group(normal SGC-7901 cells),esomeprazole acts on the experimental group(SGC-7901 cells transfected with miRNA-22-3p inhibitor)for 48 hours,the difference in cell proliferation activity was statistically significant(P<0.05).Conclusions1.Esomeprazole significantly affects the miRNA expression profile of gastric cancer cells,and the biological mechanism may be related to cAMP,MAPK,cancer?cGMP-PKG and other signaling pathway.2.Esomeprazole can up-regulate the expression of miRNA-22-3P,and inhibiting the expression of miRNA-22-3P can reduce the inhibitory effect of esomeprazole on the proliferation of gastric SGC-7901 cells. |
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