Fig. 5

NAC and GSH counteracted the therapeutic effect of Sorafenib in human HCC both in vitro and in vivo. a Quantification of the relative numbers of MHCC-97L and Hep3B cells treated with Sorafenib (5 µM), combined Sorafenib and NAC (1 mM), and combined Sorafenib and GSH (100 µM). Data were normalized and compared to the control. Results represent 3 independent experiments. b ROS assay with CM-H2DCFDA staining. Data represent the intensities of FITC normalized with control. c Annexin V assay for quantification of apoptotic cell population in MHCC-97L and Hep3B cells subjected to Sorafenib (5 µM), combined Sorafenib and NAC, and combined Sorafenib and GSH treatments. The data shown are representative of three independent experiments. d A schematic summary of Sorafenib treatment in vivo. e Bioluminescent images of mice bearing orthotopic MHCC-97L cell-derived xenograft tumors with treatment of Sorafenib (10 mg/kg), and combined Sorafenib and NAC treatment at both low and high doses. f Liver tumors with mice treated with Sorafenib and combined Sorafenib and NAC at both low and high doses and their tumor masses. (g) Bioluminescent images of lung tissues and quantification of their bioluminescent intensities. h Representative flow cytometry histogram and quantification of intra-tumoral ROS levels in the indicated groups. i Bioluminescent images of mice bearing orthotopic MHCC-97L cell-derived xenograft tumors with treatment of Sorafenib (10 mg/kg), and combined Sorafenib and GSH at both low and high doses. j Liver tumors subjected to Sorafenib (10 mg/kg), and combined Sorafenib and GSH treatment at both low and high doses and their tumor masses. k Representative flow cytometry histogram and quantification of ROS levels in vehicle control, Sorafenib alone and combination groups. Data are expressed as mean ± SD. *P < 0.05, ** P < 0.01, and ***P < 0.001 vs. Control or Sorafenib; one-way ANOVA followed by Dunnett comparison test