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Figures
- Figure 1.
Short-term culture of fragments of hepatocellular carcinoma. A: Histological examination of fragments prepared from Hep3B tumours grown as xenografts in nude mice and maintained in culture for 0 to 72 h (magnification, ×20). Note that the architecture and the heterogeneous composition of the tumours are preserved for up to 72 h. B: Analysis of tumor viability by Lactate dehydrogenase (LDH) release assay. Results are expressed as a percentage of total LDH for three independent Hep3B tumors. For each tumor, average values were obtained from four fragments analyzed at the indicated time points. C: Experimental verification of the kinase-blocking activity of sorafenib in Hep3B tumour fragments. Tumor fragments were exposed to increasing concentrations of sorafenib (0 to 10 μM) for 48 h, and protein extracts were analyzed by immunoblotting to detect Extracellular signal-regulated kinase (ERK) phosphorylation.
- Figure 2.
Heterogeneous impact of sorafenib on the ERK kinase in human hepatocellular carcinoma fragments. Tumour fragments were incubated for 48 h with increasing concentrations of sorafenib (0 to 10 μM), either alone or with erlotinib (1 μM) and processed for the analysis of ERK phosphorylation by immunoblotting. Representative immunoblots are shown for the three types of responses that were observed, i.e. tumors that are either sensitive to the inhibitory action of sorafenib (patient #6), poorly-sensitive to sorafenib (patient #5), or present a paradoxy activation of ERK phosphorylation (patient #1).
- Figure 3.
Quantification of the impact of sorafenib on ERK phosphorylation in human hepatocellular carcinoma fragments. Quantification of the ERK phosphorylation level is presented for each patient under the same experimental conditions as in Figure 2. The results are average measurements calculated from three fragments of the same tumour exposed to the indicated concentrations of sorafenib (0–10 μM), alone, and in association with erlotinib (1 μM). *p<0.05 compared to the condition without sorafenib (S0); #p<0.05 compared to equivalent conditions without erlotinib.
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