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.

(A) 为检测血管内皮生长因子受体2（VEGFR2）、血小板衍生生长因子受体β（PDGFRβ）及c-Raf的mRNA水平，以SK-Hep-1、Hep3B、Huh-7及HepG2细胞系来源的互补DNA（cDNA）为模板，开展实时定量逆转录聚合酶链反应（real-time quantitative RT-PCR）实验。以各基因的2−ΔCt值绘制柱状图展示基因表达水平，并以星号（*）标记各组细胞间基因表达的显著性差异。(B、C) 采用WST-1细胞活力检测法评估肝细胞癌（HCC）细胞系对索拉非尼处理的敏感性，以星号（*）标记SK-Hep-1与其余细胞间的显著性差异。(B) 将SK-Hep-1、Hep3B、Huh-7及HepG2细胞置于不同浓度的索拉非尼培养基中孵育24小时。(C) 将SK-Hep-1、Hep3B、Huh-7及HepG2细胞用2.5 μM索拉非尼处理不同时长。在含索拉非尼的培养基中孵育结束后，向细胞中加入WST-1试剂孵育2小时，随后检测细胞活力。针对浓度依赖性与时间依赖性数据，最终细胞存活百分率分别以溶剂对照组与0小时组的检测值为参照计算得到。