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Objective To investigate the synergistic effect of dichloroacetate (DCA) and sorafenib on the inhibition of proliferation in hepatocellular carcinoma (HCC) cells. Methods Hep3B cells were treated respectively, with DMSO (control), DCA (5 mmol/L), sorafenib (10 μmol/L), and DCA (5 mmol/L) combined with sorafenib (10 μmol/L) for 24 h, then the cells were stained with crystal violet and observed under a microscope for cell morphology. CCK-8 assay was used to detect the changes of cell proliferation, and flow cytometry was employed to measure cell apoptosis. Then, Western blotting was utilized to detect the protein expression of apoptosis-related protein PARP and p-JNK, and the production of reactive oxygen species (ROS) was measured by ROS detection kits, Finally, flow cytometry was performed to detect the changes of apoptosis after addition of anti-oxidant, NAC and JNK inhibitor, SP600125. Results DCA combined with sorafenib significantly changed the cell morphology and killed the cells. Compared with DCA alone or sorafenib alone, DCA combined sorafenib significantly enhanced the inhibitory effect on the proliferation in Hep3B cells (P < 0.05), increased the cleavage of PARP and the phosphorylation of JNK. Moreover, the combination induced the production of intracellular ROS (P < 0.05). Co-treatment of antioxidant NAC resulted in significant inhibition of elevated JNK phosphorylation and inhibitory effect on proliferation induced by DCA combined sorafenib in Hep3B cells. Conclusion Combination of DCA and sorafenib significantly inhibits proliferation in Hep3B cells, which may be related with the activation of ROS/JNK signal pathway.