Research Paper Volume 15, Issue 12 pp 5412—5425

Tumor-suppressive function and mechanism of miR-873-5p in glioblastoma: evidence based on bioinformatics analysis and experimental validation

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Figure 1. miR-873-5p, downregulated in GBM tissues or cells, targets HMOX1 and reduces its expression. (A) A volcano plot of the differentially expressed genes in GBM and normal samples from the GSE103229 dataset. Red dots indicate significantly up-regulated genes, and blue dots indicate significantly downregulated genes. The top hsa-miR-873-5p with the most significant differential expression (the lowest p value) was labeled. (B) Venn diagram of the downstream genes of miR-873-5p predicted using the miRDB, TargetScan, and mirDIP databases with the significantly up-regulated genes in GBM samples from the GSE104267 dataset. (C) HMOX1 expression in normal brain tissue samples and GBM samples analyzed by the GEPIA database. The red box represents the GBM tissue samples (n = 163), and the gray box represents the normal brain tissue samples (n = 207). (D) Binding sites of miR-873-5p in HMOX1 mRNA predicted by the miRDB database. (E) Expression of miR-873-5p and HMOX1 in tumor tissues of GBM patients (n = 20) and normal brain tissues (n = 20) measured by RT-qPCR. (F) Expression of miR-873-5p and HMOX1 in human normal brain glial cell line HEB and GBM cell lines T98G, U251, LN-229, and A172 determined by RT-qPCR. (G) Binding of miR-873-5p to HMOX1 validated by dual-luciferase reporter assay. (H) Expression of miR-873-5p in T98G cells transfected with miR-873-5p mimic or miR-873-5p inhibitor determined by RT-qPCR. (I) Western blot of HMOX1 protein in T98G cells transfected with miR-873-5p mimic or miR-873-5p inhibitor. *p < 0.05, compared to normal brain tissues, HEB cells or NC mimic-transfected T98G cells. #p < 0.05, compared to NC inhibitor-transfected T98G cells.