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. 2020 Apr 27;12(5):1081. doi: 10.3390/cancers12051081

Figure 6.

Figure 6

MiR-155-5p decreased RNF123 expression and gave a poor prognosis in GB patients. (A) TCGA analysis of a merged cohort of low-grade glioma and GB for copy number variations and mutations. The frequency of RNF123 alteration is 0.8% of a total of 1084 patients. (B) Correlation analysis of miR-155-5p and RNF123 expression using TCGA dataset from GB tumors (n = 145; Pearson r = −0.277, p = 0.0007). (C) TCGA database analysis of miR-155 in GB tissue (n = 202) compared to normal brain tissue (n = 11) (t-test, *** p < 0.001). (D) Rembrandt database analysis of miR-155 expression in GB tissue (n = 214) compared to normal brain tissue (n = 21) (t-test, *** p < 0.001). (E) TCGA database analysis of RNA-sequencing data for miR-155 in IDH WT (n = 145) or mutated (n = 8) GB tissue compared to normal brain tissue (n = 5) (one-way ANOVA, *** p < 0.001, NS = non-significant). (F) LN18, A172, and HS683 cell lines were transfected with pre-miR-155-5p (miR-155-5p-OV) or miR control (miR-Ctrl) and RNF123 expression was quantified by Western blot. (G) miR-155-5p sequence aligned with human RNF123 WT 3′-UTR (WT) and RNF123 Mutant 3′-UTR (Mut) sequences. (H) Luciferase reporter activity assay to determine the effect of miR-155-5p on 3′-UTR of RNF123 using human RNF123 3′-UTR (WT) and RNF123 Mutant 3′-UTR (Mut) sequences cloned in RenSP vector (t-test, NS = non-significant, *** p < 0.001). (I) Percentage of invasion of LN18 cell lines with miR-155-5p-OV, RNF123-OV, or both compared to control cell lines (one-way ANOVA, * p < 0.05, NS = non-significant). (J) Correlation analysis of miR-155-5p and SERPINE1 expression using TCGA dataset from GB tumors (n = 145; Pearson r = 0.368, p < 0.0001). (K) GB patients from the TCGA database were split into low (n = 90) and high (n = 90) miR-155-5p expression and analyzed for RNF123 expression (t-test, * p < 0.05). (L) Kaplan–Meier curves for the OS of GB patients expressing low (n = 87) versus high (n = 87) miR-155-5p (log-rank test, p = 0.024). (M) Dot plot to determine miR-155-5p expression in pre-operative plasma from GB patients (n = 19) and plasma of healthy controls (n = 46) (t-test, ** p = 0.024). Error bars represent mean ± SD from replicates (n = 3).