title: Enhancement of cytotoxic and pro-apoptotic effects of 2-aminophenoxazine-3-one             on the rat hepatocellular carcinoma cell line dRLh-84, the human hepatocellular             carcinoma cell line HepG2, and the rat normal hepatocellular cell line RLN-10             in combination with 2-deoxy-D-glucose.
abstract: The cytotoxic and pro-apoptotic effects of a single dose of 2-aminophenoxazine-3-one             (Phx-3) or 2-deoxyglucose (2-DG) or of a combined dose of Phx-3 and 2-DG were             studied in the rat hepatocellular carcinoma cell line dRLh-84, the human hepatocellular             carcinoma cell line HepG2 and the rat normal hepatocellular cell line RLN-10.             The number of viable cells decreased in a dose-dependent manner, when dRLh-84,             HepG2 or RLN-10 cells were treated with 2-DG (0.5-20 mM) or Phx-3 (1-50 µM) alone             at 37˚C for 48 h. When these cells were treated with 10 mM 2-DG and different             concentrations of Phx-3, the number of viable cells decreased dose-dependently             and in an additive manner for these agents. A single dose of 2 or 10 µM Phx-3             induced apoptotic morphology characterized by nuclear condensation and cell shrinkage             in dRLh-84, HepG2 and RLN-10 cells, while a single dose of 10 mM 2-DG did not.             When Phx-3 (2 or 10 µM) treatment was combined with 2-DG (10 mM) treatment in             these three cell lines, the cells with apoptotic morphology increased extensively,             which was confirmed by flow cytometric analysis. In addition, autophagic morphology             characterized by cytosolic vacuole formation was significantly increased in the             hepatocellular carcinoma cell lines dRLh-84 and HepG2 but not in the normal hepatocellular             cell line RLN-10 after a single dose of Phx-3 or 2-DG or a combined dose of Phx-3             and 2-DG. Furthermore, when dRLh-84 and HepG2 cells were treated with Phx-3 alone             or a combined dose of Phx-3 and 2-DG, depolarization of the mitochondria was extensive,             but that of the normal cell line RLN-10 was not. These results may imply that             the mechanism for the apoptosis of hepatocellular carcinoma cells caused by Phx-3             alone or a combined dose of Phx-3 and 2-DG differs from that of the normal cell             line RLN-10. The present results demonstrate that Phx-3 alone may be beneficial             for targeting liver cancer and that its anticancer activity may be enhanced by             2-DG. However, a combined dose of Phx-3 and 2-DG may exert adverse effects on             normal liver cells, as evidenced by the cytotoxic and pro-apoptotic effects of             the combined treatment in the rat normal hepatocellular cell line RLN-10.
authors.affiliation: Fourth Department of Surgery, Tokyo Medical University, Ibaraki                     Medical Center, 3-20-1 Chuo, Inashiki, Ibaraki 300-0395, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Takemura
authors.forename: Xiao-Fang
authors.initials: XF
authors.lastname: Che
authors.forename: Takafumi
authors.initials: T
authors.lastname: Tabuchi
authors.forename: Shota
authors.initials: S
authors.lastname: Moriya
authors.forename: Keisuke
authors.initials: K
authors.lastname: Miyazawa
authors.forename: Akio
authors.initials: A
authors.lastname: Tomoda
mesh_terms: D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000473:pathology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003847:Deoxyglucose / Q000627:therapeutic use / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000473:pathology; D008114:Liver Neoplasms, Experimental / Q000188:drug therapy / Q000473:pathology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010078:Oxazines / Q000627:therapeutic use / Q000633:toxicity*; D051381:Rats
chemical_list: D010078:Oxazines; C005512:3-aminophenoxazone; D003847:Deoxyglucose