Figure 5.

Growth inhibition by the anti-sense oligonucleotide targeting SKP2 mRNA. ACC-LC-172 cells, exhibiting amplification and overexpression of SKP2, were treated with an anti-sense oligonucleotide targeting SKP2 (AS) or a scramble oligonucleotide (SC) by using Oligofectamine. A: Levels of SKP2 determined by Western blotting. Cells were treated with 200 nmol/L of oligonucleotides (anti-sense or scrambled) or Oligofectamine alone, and then harvested at 48 hours after transfection. Untreated cells were maintained under identical experimental conditions. Six μg of whole cell lysate were separated by polyacrylamide gel electrophoresis and analyzed by immunoblotting. B and C: The effects of anti-sense oligonucleotide on the viability of ACC-LC-172 cells. Cells were treated with the indicated concentrations of oligonucleotides (anti-sense or scrambled) and cell viability was determined by MTT assay at 4 days after transfection (B). Cells were treated with 200 nmol/L of oligonucleotides (anti-sense or scrambled) and cell viability was determined at the indicated times after transfection (C). The percentage was calculated against the absorbance of the control cells treated with Oligofectamine alone. The values were shown on the plot represent the means ± SD of four independent experiments.