Figure 3.

Knockdown of MOR attenuates the proliferation and migration of HCC cells. (A and B) HepG2 and Huh7 cells were transfected with control or MOR siRNA, and forty-eight hours later, total RNA and whole cell lysates were extracted and subjected to real-time PCR and western blot analysis for MOR expression, respectively. MOR was effectively silenced, as shown by the mRNA and protein levels of the two cell lines (***P < 0.001). (C) Recovered MOR expression in HepG2 and Huh7 cells after transfection with the pCI-neo-MOR plasmid was analyzed by western blot. (D) The colony formation ability of MOR-silenced HepG2 and Huh7 cells was inhibited, while overexpressing MOR using the pCI-neo-MOR plasmid reversed the inhibition effects (***P < 0.001; ****P < 0.0001). (E) The proliferation of MOR-silenced HepG2 and Huh7 cells was suppressed, and regaining MOR rescued proliferation (**P < 0.01; ***P < 0.001; ****P < 0.0001). (F) Wound healing assays showed that the migration ability of HepG2 and Huh7 cells was attenuated by MOR knockdown, and transfection with pCI-neo-MOR rescued migration (scale bars, 100 μm; *P < 0.05, **P < 0.01; ***P < 0.001). (G) Transwell assays showed that the migration ability of HepG2 and Huh7 cells was attenuated by MOR knockdown, and transfection with pCI-neo-MOR also rescued migration (scale bars, 100 μm; *P <0.05, **P < 0.01; ***P < 0.001). All experiments were repeated three times independently. See Supplementary File Raw Blot Images (Figure 3B and 3C) for original blot images. Data were presented as the mean ± SEM. One-way ANOVA methods were used for statistical analysis.