Transcriptional Regulation of Fucosyltransferase 1 Gene Expression in Colon Cancer Cells
Figure 3
Site-directed mutagenesis on FUT1 promoter and effect of DN-Elk-1 overexpression on expression of FUT1 mRNA in DLD-1 cells. (a) Mutagenesis in the region −90 to −81 of the FUT1 promoter. 10 μg of the wild (pGL4.11/−190_+1_wild) and mutant (pGL4.11/−190_+1_mut) constructs and 0.5 μg of pRL-CMV were co-transfected into DLD-1 cells and luciferase activity was determined 24 h aftertransfection. In each case, firefly luciferase activity was normalized to Renilla luciferase activity to correct for differences in transfection efficiency. The relative activities of luciferase are normalized to the activities of pGL4.11/−190_+1_wild as 100. The results obtained in three independent experiments are expressed as mean ± SD. Significant differences () compared with the equivalent wild-type reporter construct are indicated by asterisks. (b) FUT1 mRNA expression in DN-Elk-1-transfected cells. After transfection with 5, 10, and 20 μg DN-Elk-1 expression vector, FUT1 mRNA expression levels in DLD-1 cells were determined by RT-PCR. The results were obtained in three independent experiments and data represent mean ± SD. Significant differences () compared with non-transfected cells are indicated by asterisks.
