Figure 4

ANXA2 and KRAS are upregulated in PDAC tumors and direct targets of miR-206. (a) Schematic representation of the ANXA2 and KRAS mRNA with the predicted target sites for miR-206. (b) Luciferase reporter assay in PANC-1 cells transfected with vectors containing either the wild-type or the mutated 3′ UTRs of ANXA2 or KRAS and miR-206 mimic or negative control. Experiments were performed with six replicates and data are shown as mean±s.d. (c) ANXA2 and KRAS mRNA levels after transfection of PANC-1 or PANC10.05 cells with miR-206 mimic or control. Total RNA was extracted 48 h after transfection and mRNA levels were determined by qRT-PCR. Experiments were performed with three replicates and in three independent experiments. Data are shown as mean±s.d. (d) Total cell lysates of PANC-1 or PANC10.05 were analyzed for ANXA2 protein levels 48 h after transfection with miR-206 mimic or control by western blot. (e) Expression of ANXA2 or KRAS in normal (n=7) and tumor (n=25) pancreatic tissues (GSE32688 data set). Data represent log2 expression values. Horizontal line represents the median (Whiskers: Tukey). Data were analyzed using two-tailed unpaired t-test, *P<0.05, **P<0.01, ***P<0.001. (f) Correlation analysis of miR-206 and ANXA2 or KRAS mRNA expression in PDAC patients with each data point representing an individual sample and correlation coefficient (r) indicated (Pearson correlation; GSE32688 data set). Data represent log2 expression values.