Figure 1.

TKT silencing inhibited cell migration and invasion but minimal effect on cell growth. (A) Silencing TKT expression by shRNA. Tow plasmid clones of TKT-shRNA sh1 and sh2 were constructed, and transfected into CE48T/VGH cells for 1 to 3 days. The cellular proteins were extracted for determination of TKT expression by western blot method. Actin protein level was determined as internal control. (B) TKT silencing has minimal effect on cell growth. Total of 5 × 10⁵ cells were seeded in a 10-mm dish plate. After transfection with TKT-shRNA or the vector plasmids, cells were continuously cultured for up to 4 days. Cell numbers were counted every 24 hours. (C) TKT silencing inhibited cell migration. After transfection with TKT-shRNA or the vector plasmids, cells were seeded in the upper chamber of the Transwell polycarbonate chamber. After 12 or 24 hours, the cells migrated to the lower chamber were fixed and stained with 0.5% crystal violet and photography. (D) TKT silencing suppressed cell invasion. After transfection with TKT-shRNA or the vector plasmids, cells were seeded in the 24-well plate coated with Matrigel in a Millicell invasion chamber. The numbers of cells invading through the Matrigel to the lower chamber were determined daily for 4 days. All experiments were performed in triplicate.