Effect of pathway inhibitors on AFP mRNA in HepG2/C3A and PLC/PRF/5 cells.

<p>The cells were seeded in a six-well plate at a density of 10⁴ cells/well. At 80% confluence, the cells were treated with inhibitors of PI3k/mTOR/Akt and costunolide for 48 h. The cell RNA was then extracted for RT-PCR, and this step was followed by cDNA amplification. The cDNA was then run in a 2% agarose gel and visualized by SYBR Safe staining. The relative mRNA levels were obtained using the Gel Analyzer software. Each experiment was repeated at least three different times.