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Archive > Volume 29 Issue 12 > 2022,29(12):1094-1100. DOI:10.3872/j.issn.1007-385X.2022.12.005 Prev Next
Basic Research
Screening and validation of markers related to diagnosis and prognosis assessment of gastric cancer based on miRNA-mRNA network and their potential molecular mechanisms
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Abstract:
Objective: To explore the role of miR-1-3p, a gastric cancer-related biomarker, on gastric cancer cell proliferation and its mechanism by bioinformatics approach. Methods: Transcriptomic data from gastric cancer (n=375) and paraneoplastic tissues (n=45) in the TCGA database were collected to construct a gastric cancer-specific mRNA-miRNA network, screen potential miRNA-like markers, predict the downstream target genes of the markers and analyze their functions using TargetScan. Human normal gastric epithelial cell and gastric cancer cell lines have been selected, and their miR-1-3p and myocardin (MYOCD expression was detected by qPCR. The miR-1-3p mimics were transfected into GES-1, AGS, and MKN45 cells using lipofectamine 2000. The proliferation ability of the cells was determined by CCK-8 assay, the expression of MYOCD was measured by WB assay, and the targeting relationship between miR-1-3p and MYOCD was verified by dual luciferase reporter assay. Results: Differentially expressed 259 miRNAs and 7 545 mRNAs were obtained by database data analysis to construct a regulatory network of gastric cancer-specific mRNA-miRNAs. Analysis of vulnerability structure in the network identified miR-1-3p as a potential gastric cancer marker, and in-depth analysis revealed its significance for the diagnosis and prognostic assessment of gastric cancer, while predicting MYOCD as its downstream target. Cellular assays showed that miR-1-3p was lowly expressed in gastric cancer cells (P<0.05); overexpression of miR-1-3p inhibited the proliferation ability of AGS and MKN-45 in gastric cancer cells (P<0.05 or P<0.01); and inhibited the expression of MYOCD (P<0.01); miR-1-3p was predicted to be associated with two binding sites in the 3'UTR region by the TargetScan database, and dual luciferase reporter assays showed that high expression of miR-1-3p at one of the wild-type predicted sites significantly inhibited MYOCD expression (P<0.01). Conclusion:miR-1-3p may be a potential diagnostic and prognostic-related marker for gastric cancer, and miR-1-3p may affect gastric cancer by targeting MYOCD.
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