Figure 1From: Novel synergistic antitumor effects of rapamycin with bortezomib on hepatocellular carcinoma cells and orthotopic tumor model Combined treatment with rapamycin and bortezomib inhibited cell proliferation and enhanced cell apoptosis. (A) Proliferation of HCCLM3 cells was evaluated by using Cell Counting Kit-8 (CCK-8) at indicated time points. (B) HCCLM3 cells were treated with rapamycin (10 ng/ml), bortezomib (100 nM) or both agents. ModFit software analysis of flow cytometry histograms revealed that rapamycin treatment resulted in cell cycle arrest at G1-S phase. Bortezomib significantly increased in the percentage of cells in the G2/M phase. When the two agents were combined, no significant difference in cell cycle distribution was observed compared with bortezomib alone. (C) HCCLM3 cells were treated with rapamycin (10 ng/ml), bortezomib (100 nM) or the combination for 24 or 48 h and stained with Hoechst 33342, the apoptotic nuclear changes (arrows) were examined by fluorescence microscopy (magnification, ×400). (D, E) The quantification of apoptotic cells induced by rapamycin and bortezomib was further confirmed by flow cytometry analysis. The sub-G1 contents were designed as apoptotic cells. * P < 0.01, versus control group; ** P < 0.001, versus control group; # P < 0.05, versus bortezomib treatment group at 24 h; †P < 0.01, versus bortezomib treatment group at 48 h. (F) The cells were treated with various concentrations of the drugs for 24 h, and then evaluated by using Cell Counting Kit-8. †P < 0.01, versus bortezomib treatment group.Back to article page