Figure 3
FUT8 is the specific target of miR-198 in colorectal cancer cells and has a negative correlation with miR-198 expression in CRC tissues.
SW1116 and HCT116 cells were stably transfected with a miR-198 overexpression plasmid or control negative plasmid for subsequent analysis. Data in all statistical plots represent means ± SD. (A) Overlap analysis revealed FUT8 in all the three independent online databases: PicTar, TargetScan and miRDB. (B) Representative Western blot and analyzed data showing that miR-198 overexpression significantly decreases total FUT8 expression in SW1116 and HCT116 cells. Statistical analysis is based on three independent experiments which have been run under the same experimental conditions and representative Western blot images are shown. (C) Luciferase reporter assays suggest that miR-198 inhibits FUT8 promoter activity but not the mutant sequence devoid of the putative binding sites. The luciferase activity is calculated as the ratio of firefly (reporter)/renilla (normalization) signals. (D) qRT-PCR analysis reveals a negative correlation between miR-198 and FUT8 mRNA expression in CRC tissues (P = 0.01, paired t-test).
