A Study On The The Effects Of Endostar In The The Model Of Mouse Hepatoma H22

Objective: To observe the antiangiogenic effect of homemade Recombinant humanendostatin(Endostar) in the model of mouse hepatoma H22,and have a preliminaryprobe into its mechanism by observing the tumor growth.Method:1.The establishment of the model of mouse hepatoma H22: After the-80℃frozen-stored H22cells recovery,injected into the left armpit of kunming mice,7dayslater, tumor generated obvious.Took tumor tissue under the aseptic condition,adjustedthe tumor cell density of liquor to1×107cells/ml,injector tumor suspension liquid0.2ml and injected into the subcutaneous of right armpit of324-5weeks old malekunming mice,established the mice model.Then the mice were randomly devided intoA、B、C、D groups，each group of8mice.2.Drug administration:administered drug to A group from the first day, B group from the5th day,C group from the9th day. Method of administration:Endostar abdominalinjection,3mg/Kg,inoculated tumor to4groups simultaneity from the5th day,the Dgroup only accepted tumor but did not accept drug.killed all mice in the18th day,Leavingthe tumor tissue,standby application.3.sample preparation:Took blood from the eyes,then killed and dissected the mice,weighing tumor.all specimens were fixed with10%formalin,immersed in the wax,cuted into5μm slices,xylene dewaxing, gradient alcohol to hydration;1%methanol hydrogen peroxidesolution soaking;0.125%of trypsin digestion, nonimmunologic animal serum closed,fight CD34monoclonal antibody specimens, incubation60minutes at roomtemperature; HuaEr biotin resistance,3710minutes streptavidin-which peroxidase at10minutes incubation solution, control show color by the DAB microscopic,sappanwood counterstain, neutral gum sealed piece.4.Outcome measures:from the9th day of the experiment,measured the tumor of allmice everyday to count the volume,it is equal to major axis times transverse diametertwo squared over2.Calculated the tumor microvascular density. 5.Statistical analysis:Analyzed wether there was statistically significant between thetumor volume and microvessel density of drug abdominal injection group in the18thday of the experiment and that of the control group.Using SPSS17.0statisticalsoftware for statistical data analysis,using single factor analysis of variance for theariance inspection of all data, further the LSD-t test was used to pairwise comparison.Results:The18th day of the experiment: there was statistically significant betweenthe tumor volume and microvessel density of drug abdominal injection group(B) inthe day of inoculated tumor and that of the control group(D)(P<0.05). There was nosignificant differences between the tumor volume and microvessel density of drugadministration group before (A)and after (C)the day of inoculated tumor and that ofthe control group(P>0.05).Conclusion:The application of Endostar alone shows anti-tumor effect,the best timeof drug administration is the time of inoculated tumor.