Effect of telomerase, mTOR and PKC inhibitors on HepG2/C3A and PLC/PRF/5 cell migration.

<p>The migration of HepG2/C3A and <b>PLC/PRF/5</b> cells was assayed using the Boyden chamber technique in a 24-well plate. According to the manufacturer’s recommendations, 10⁶ cells in DMEM without serum were suspended in the upper chamber of each well; in the lower chamber, DMEM + 10% FBS was added as a chemo-attractant solution. After 24 h of treatment, the cells that migrated to the lower chamber were stained using a staining solution that fixes and stains the cell walls and cytoplasmic membranes of any cell remaining on the insert. The stain is instantly dissolved once the kit extraction solution is added. The solution was then transferred to a 96-well microtiter plate, and the absorbance was measured at 560 nm using a plate reader. (A) Costunolide (10 μM), BIBR (10 μM), rapamycin (200 nM) and SPC0213 (5 μM) had a significant effect on HepG2/C3A cell migration after treatment for 24 h. (B) Costunolide (50 μM), BIBR (50 μM) and SPC0213 (5 μM) had a significant effect on PLC/PRF/5 cell migration after treatment for 24 h. Each value represents the mean of three assays. Each value was normalized to its own control, which was set to 100%. Each point represents the mean ± SD from all of the experiments. *P<0.01 versus untreated cells. **P<0.005 versus untreated cells.