Fig. 3.

Integrative analysis of miRNA and mRNA expression profiles of the subtypes. (A) Supervised analysis of miRNA expression profiles of Cluster 1 and Cluster 2 of the initial 100 cases identified 10 differentially expressed miRNAs (statistical significance of each difference indicated in the bar graph). (B) ACVR1 mRNA expression was significantly higher in Cluster 1 (P = 0.02). (C) The ACVR1 3′-UTR has one binding site with miR-148a, as predicted from TargetScan. MHCC97H cells transfected with pmirGLO-ACVR1 and pmirGLO-ACVR1-Mut reporters, together with a miR-148a mimic or mimic negative control, miR-148a overexpression suppressed the activity of luciferase in the wild-type (P = 0.021) but not in mutant type (P = 0.596). (D) ACVR1 expression was negatively correlated with miR-148a expression (r = −0.22, P = 0.027). The 100 HCC patients were categorized into three groups based on miRNA-148a expression (low, expression <1; middle, 1 ≤ expression ≤2; high, expression >2), and overall survival in these three groups differed significantly (P = 0.03). (E) MiR-148a regulated Wnt signaling targets by down-regulating ACVR1 expression. Up-regulated genes in Cluster 1 are indicated in red, and the intensity of the red indicates the relative log2 ratio as shown in the color bar. The cyan lines and blockers represent the direction of the miRNAs, and + and − indicate positive and negative regulation of gene expression, respectively. Gray lines represent known physical interaction between genes connected.