Fig. 6

Clinical implications of miR-2355-5p. (A) Volcano plot summarizing the RNA expression profiles of ccRCC patients obtained from the TCGA-KIRC (top) and CPTAC3 (bottom) databases (TCGA-KIRC: N = 72, CPTAC3: N = 149). RNAseq analysis was performed similarly to miRNAseq previously described. (B) Left: Venn diagram comparing enriched protein transcripts and significantly downregulated genes in ccRCC tumors from TCGA-KIRC and CPTAC3 cohorts. Right: Venn diagram comparing selected protein transcripts from the left diagram (marked in green) and predicted miR-2355-5p targets from prediction software (miRDB, TargetScan, miRWalk). C-D RT-qPCR quantification of selected potential target genes in 786-0 (B) and A498 (C) Cr.2355 models compared to appropriate controls (N = 4–6). E. RT-qPCR quantification of selected genes in ccRCC primary tissue compared to normal tissue (N = 13). F Tumor growth of the 786-0 Cr.2355 xenograft mouse model (N = 4–8). G Image illustration of extracted xenograft tumors H-I RT-qPCR quantification of miR-2355-5p (H) and selected targets (I) in 786-0 Cr.2355 mouse tumors (N = 4–8). J Tumor growth of the 786-0 + + 2355 xenograft mouse model (N = 6–7). K-L. RT-qPCR quantification of miR-2355-5p (K) and selected targets (L) in 786-0 + + 2355 mouse tumors (N = 6–7). MiRNA and gene expression was normalized with RNU44 (H, J), RPLPO (C, D,K, L), or RPLPO + HPRT1 (E) expression. The data are presented as the Mean ± SEM. Statistical analysis was performed using two-tailed unpaired (C, D, H, J, K, L) or paired (E) Student’s t-tests or two-way ANOVA with Dunnett’s multiple comparison test compared to 786-0 Cr.CTRL (F) and 786-0 + + CTR: (I). (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)