Figure 5.

MicroRNAs (miRNAs) that regulate HULC expression in pancreatic ductal adenocarcinoma (PDAC) cells. Panc‐1 cells were treated with 0 (control) or 10 ng/mL transforming growth factor (TGF)‐β. After incubation for 72 h, RNA was isolated, and expression profiling of 2555 miRNAs was carried out. Expression of 1719 miRNAs was detected in Panc‐1 cells. A, Scatter plot of the microarray intensities of TGF‐β‐treated Panc‐1 cells against those of control cells. B, Waterfall plot of the 187 miRNAs whose expression was decreased <0.87‐fold in Panc‐1 cells treated with TGF‐β. C, MicroRNA (MiR)‐133b was predicted to target HULC by bioinformatics analysis using miRNA.org. D, RNA was extracted and miR‐133b expression was assessed by quantitative RT‐PCR in PDAC cells after incubation with 10 ng/mL TGF‐β for 72 h. MiR‐133b expression relative to the controls is shown. Gene expression was normalized to that of RNU6B. E, Panc‐1 and MiaPaCa‐2 cells were transfected with 12.5 nmol/L miR‐133b or control mimic, and after 24 h, were cotransfected with 2.0 µg HULC firefly luciferase reporter vector or empty vector and 0.1 μg Renilla luciferase reporter pRL‐SV40. After a further 24 h, relative firefly luciferase activity was measured and normalized to Renilla activity. Bars are means ± SEM of 3 independent experiments. *P < .05