| Objective: To study the effects of simvastatin onproliferation and apoptosis of liver cancer SMMC-7721 andH22.Methods: 1,cellular experiment SMMC-7721 wasgrown in RPMI1640 medium supplemented with 8%heat-inactived fetal calf serum. MTT assay: After treatedSMMC-7721 cell with simvastatin 24-72 hours, the proliferationof SMMC-7721 were examined with MTT test . After treatedSMMC-7721 cell with simvastatin 48 hours ,the distribution ofcell cycle and factors associated such as bcl-2,cdk-2,NF-κB,p21Ras were examined by FCM. To examine the interactionbetween simvastatin and priarubicin, The interaction index forcombinations of the two drugs was computed. When theinteraction index is less than 1.0, Synergy occurs. 2 animalexperiment H22 cell was grown in the abdominal cavity ofmice, on the inoculation day, the tumor cells were harvestedfrom the abdominal cavity and washed with D-hank's, and then1×106 H22 cells in 200 μl of medium were injected into theback of the KM mice . Mice were randomly devide into fourgroups and gastro-gavaged with simvastatin 0,10,20,40mg/d/kg starting at day 6 after inoculation of tumor cells after thetumor nodules became visible in the back. Local tumor growthwas determined by measuring back diameter every other day tillthe day when the first mice died, starting with the first day oftreatment (day 7 after inoculation of tumor cells -initial tumorvolume). Tumor volume and Tumor growth delay werecalculated to test whether simvastatin can inhabit tumour growththrough animal experiment .survival was computer to testwhether simvastatin can make prolongation of the survival.Result 1,simvastatin could inhabit the proliferation ofSMMC-7721 by a does and time depent way, up to 60%inhabition at 20μmol/L and 72hr . simvastatin was dissolved byethanol and we use 0.1% ethanol(the highest concentrationethanol of simvastatin) as contral concentration to get ride of theeffect of ethanol .showing that 0.1% ethanol have no effect onproliferation of SMMC-7721 2,cell cycle distribution show5,10,20μmol/L simvastatin can induce SMMC-7721apoptosis,after treated SMMC-7721 cell with simvastatin 48hours .The apoptosis index was 3.89%,5.96%,11.13%,separately.The percentage of G1 in cell cycle have atendency to increase .3 ,To determine whether the proliferationinhabition and apoptosis accompanying simvastatin exposurewas associated with changes in the expression of p21Ras,bcl-2,cdk2,NF-κB, FCM was performed on SMMC-7721 afterexposure to 0,5,10,20μmol/L simvastatin 48 hours .The FIvalue was used to represent the expression ofp21Ras,bcl-2,NF-κB ,cdk2.(1) The FI value of p21Ras were 1,0.84,0.83,0.75, separately ,showing that the expression of p21Ras wasdecreased accompanying the increasing concentration ofsimvastatin. (2) The FI value of bcl-2 were 1,0.86,0.83,0.70separately ,showing that the expression of bcl-2 was decreasedaccompanying the increasing of concentration of simvastatin.(3)The FI value of NF-κB were 1 ,0.95 ,0.86 ,0.74,separately ,showing that the expression of NF-κB wasdecreased accompanying the increasing of concentration ofsimvastatin.(4) The FI value of were 1,0.94,0.91,0.81,separately ,showing that the expression of cdk2 was decreasedaccompanying the increasing of concentration of simvastatin .4,The isobolanalysis was usedt to determine whether simvastatinand priarubicin exert synergistic effects in the treated cells,SMMC-7721. According to this analysis, synergy occurs whenthe interaction index is less than 1.0. The interaction index forsimvastatin and priarubicin used in combination inSMMC-7721 was 0.71. Thus, the results of our in vitroexperiments suggest that potentiated effects of combinationtherapy with simvastatin and priarubicin may be due to thesynergistic interaction of these two drugs 5. animalexperiment results The median tumor growth time for micetreated with simvastatin 0,10,20,40mg/ d/kg when everygroup reach the volume of 4.8504cm3 were 10 days,10 days,12.5days,16 days,The median tumor growth delay were 0,2.5,6days. When the first mice died, we begin to study survival .In our experiments all mice died with developing tumors except...
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