Analysis of mRNA expression of sorafenib-target genes and sorafenib sensitivity in HCC cell lines.

(A) To examine the mRNA levels of VEGFR2, PDGFRβ, and c-Raf, real-time quantitative RT-PCR was performed with cDNA derived from SK-Hep-1, Hep3B, Huh-7 and HepG2 cell lines. Expression of the genes was plotted in bar graphs using 2−ΔCt values of each gene and the significant difference in gene expression among the cells were marked with asterisk (*). (B, C) The susceptibilities of HCC cell lines to sorafenib treatment were evaluated by WST-1 cell viability assay. The significant differences between SK-Hep-1 and the other cells were marked with asterisk (*). (B) SK-Hep-1, Hep3B, Huh-7 and HepG2 cells were incubated for 24 hours at the indicated concentrations of sorafenib. (C) SK-Hep-1, Hep3B, Huh-7 and HepG2 cells were treated with 2.5 μM of sorafenib for the indicated time. After incubation in sorafenib-containing media, WST-1 was added to the cells for 2 hours and then the cell viabilities were measured. The final cell survival (%) values were calculated relative to the value of vehicle and at 0 hours in concentration and time-dependent data, respectively.