Figure 5.

KPNA2 is validated as a direct target gene of miR-411 in ESCC cells. (A) The binding sequences for miR-411 in the wild-type KPNA2 3′-UTR. The mutant KPNA2 3′-UTR is also presented. (B) Eca109 and TE-1 cells were cotransfected with either miR-411 mimics or miR-NC and either the KPNA2-wt or KPNA2-mut reporter plasmid. The transfected cells were harvested after 48 h of incubation and then subjected to the detection of luciferase activity. *P<0.05 vs miR-NC. (C) RT-qPCR was carried out to assess KPNA2 mRNA expression in Eca109 and TE-1 cells after their transfection with miR-411 mimics or miR-NC. *P<0.05 vs miR-NC. (D) The protein expression of KPNA2 was determined by Western blotting in miR-411 mimics–transfected or miR-NC–transfected Eca109 and TE-1 cells. *P<0.05 vs miR-NC. (E) RT-qPCR analysis uncovered the expression profile of KPNA2 mRNA in the 51 pairs of ESCC and matched adjacent normal tissue samples. *P<0.05 vs normal tissue samples. (F) A negative correlation between miR-411 and KPNA2 mRNA levels in ESCC cells was demonstrated via Spearman’s correlation analysis. R²=0.3186, P<0.0001.