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Figure 2 | BMC Cancer

Figure 2

From: microRNA-141 inhibits cell proliferation and invasion and promotes apoptosis by targeting hepatocyte nuclear factor-3β in hepatocellular carcinoma cells

Figure 2

Prediction and validation of HNF-3β as the target of miR-141. (A) Schematic description of the hypothesized duplexes formed by the interactions between the HNF-3β 3′-UTR binding site and miR-141. The predicted structure of the base-paired hybrid is diagrammed. Paired bases are indicated by a black line, and G:U pairs are indicated by three dots. The predicted free energy of the hybrid is indicated. (B) Quantitative RT-PCR analysis of the relative miR-141 level in 12 pairs of HCC tissues and noncancerous tissue samples. (C) Analysis of luciferase activity. Firefly luciferase reporters containing either the wild-type (WT) or mutant (MUT) form of the human HNF-3β 3′-UTR were cotransfected into HepG2 cells with pre-miR-141 or pre-miR-control. At 24 h post-transfection, the cells were assayed using a luciferase assay kit. Firefly luciferase values were normalized to β-galactoidase activity and plotted as relative luciferase activity. For comparison, the luciferase activity in pre-miR-control-transfected cells was set as 1. (D) Quantitative RT-PCR analysis of the relative miR-141 expression level in HepG2 cells transfected with pre-miR-141 or pre-miR-control for 24 h. (E and F) Western bolt analysis of the endogenous HNF-3β protein level in HepG2 cells transfected with pre-miR-141 or pre-miR-control for 24 h. (E): representative image; (F): the result of the quantitative analysis. (G) Quantitative RT-PCR analysis of the relative miR-141 expression level in Huh7 cells transfected with pre-miR-141 or pre-miR-control for 24 h. (H and I) Western bolt analysis of the endogenous HNF-3β protein level in Huh7 cells transfected with pre-miR-141 or pre-miR-control for 24 h. H: representative image; I: the result of the quantitative analysis. (mean ± S.D.; * p < 0.05; *** p < 0.001).

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