Fig. 5.

Tetraspanin 1 (TSPAN1) is a downstream target of micro RNA (miR)-194-5p. (a) Analysis using TargetScan and miRanda revealed several miRNAs might regulate TSPAN1. (b) Human cholangiocarcinoma (CCA) tissues displayed significantly lower miR-194-5p levels than adjacent nontumor tissues did. (c) Binding site of miR-194-5p in wild-type (WT) 3′-untranslated region (UTR) of TSPAN1 and corresponding mutant type were constructed. (d) Luciferase reporter assay showed luciferase activity of HEK293T transfected with WT 3′-UTR was obviously inhibited by miR-194-5p overexpression. (e) miR-194-5p overexpression or silencing decreased or increased TSPAN1 protein in indicated cells. (f) Correlation analysis indicated an inverse correlation between miR-194-5p and TSPAN1 expression. (g) Kaplan-Meier analysis indicates that combination of high TSPAN1 and low miR-194-5p predicts poorer overall survival rate than low TSPAN1 and high miR-194-5p. All data are means ± SD of three separate experiments. ***p < 0.001 and ****p < 0.0001