Research On Antitumor Activity And Mechanism Of Curcuminderivative C086 With The Orthotopic Liver Transplantation Model

Objective: Set up xenograft tumor model and orthotopic transplantation tumor model of liver cancer and study its antitumor effects and mechanism of curcumin derivatives objects in vivo and in vitro and the combination effect of C086 with sorafenib.Methods:1 MTT assay was used for detecting the effects of proliferation inhibition of C086 and sorafenib on Hep G-2 tumor cell line. 2 H22 mice liver cancer xenograft tumor model and orthotopic transplantation tumor model were established in KM mice to evaluate the antitumor activities. 3 Hep G-2 human liver cancer xenograft tumor model and orthotopic transplantation tumor model were established in nude mice to evaluate the antitumor activities. 4 Western blot was used to analyze the related protein levels of RAS-RAF- MAPK and caspase signal pathway in Hep G-2 cell after exposing to C086 for 24 h. 5 The expression change of RAF-1, MEK, p-MEK, ERK, p-ERK in Hep G-2 nude mice orthotopic tumor was showed by immunohistochemistry and western-blot.Results:1 C086 inhibited the cell proliferation of liver cancer cell lines Hep G-2 in a dose-dependent manner with IC50 of 5.71μg·m L-1. Sorafenib had the same IC50 value as C086 at 11.06μg·m L-1. The CI value was lower than 1. 2 Results using a tumor subcutaneous of mice live cancer cell line H22 showed that 100mg/kg/d and 150mg/kg/d C086 intravenously respectively obtained tumor inhibition rates of 36.2%,43.1% without loss of body weight,while C086 200mg/kg/d intragastrically reached a highest inhibition rate of 61.4%.When using a tumor orthotopic of H22, the inhibition rate of the dosage at 200mg/kg/d changed to be 78.4%.while the inhibition rate of sorafenib at a dosage of 50mg/kg/d was just 68%. Inhibition rate had not obvious improvement after combination of C086 and sorafenib. 3 Results using a tumor subcutaneous of human liver cancer cell line Hep G-2 showed that 50 mg/kg/d,100mg/kg/d and200mg/kg/d C086 intragastrically respectively obtained tumor inhibition rates of 23.9%,27.1%,33.9%without loss of body weight,while sorafenib 50mg/kg/d reached a inhibition rate of 74.1%.And the drug combination group 84.3%. When using a tumor orthotopic of Hep G-2, the inhibition rate of the dosage of 50 mg/kg/d, 100mg/kg/d and 200mg/kg/d changed to be33.0%, 43.9%, 69.1%.while the inhibition rate of sorafenib at a dosage of 50mg/kg/d was just 66.8%. And the drug combination group 84.6%.Before the instrument deadline, every nude mouse was alive without losing weight. 4 C086 blocked RAF/MEK/ERK signaling pathway, inhibiting the phosporylation of MEK and ERK in Hep G-2 in vitro. 5 C086 inhibited RAF/MEK/ERK signaling pathway,reducing the phosporylation of RAF-1, MEK and ERK in Hep G-2liver cancer orthotopic transplantation tumor.Conclusion:1 C086 had a significant inhibition on the growth of H22 mice liver tumor and Hep G-2 liver cancer after intragastrical administration in vivo. 2 C086 shows potent efficacy against Hep G-2 liver cancer in vitro and in vivo, the mechanism of which may be based on the enhancement of Ras-Raf-MAPK proliferous pathway inhibition and caspase apoptotic induction. 3 C086 is synergistic with sorafenib on Hep G-2 in vitro.The synergism may be based on the enhancement of proliferous inhibition and apoptotic induction.