The Inhibition And Its Mechanism Of Bortezomib Which Play On H22Hepatoma

Part1The inhibitory effect and mechanism research of Proteasomeinhibitor—Bortezomib in murine hepatocellular carcinoma H22Objective: Confirm the bortezomib’s inhibition effect of hepatocellular carcinoma(H22）through the experiments in vivo study and preliminary reveal the mechanism of itsaction.Methods:bortezomib，H22cell line (murine hepatocellular carcinoma),KM mouse；Establish H22tumor model in KM mouse subcutaneously, inject bortezomib via tail vein（0.5mg/kg/4days x3）, observe the inhibition effect,reveal the mechanism viaobservation,measurement,Immunohistochemical assay.Results: Mouse model of liver cancer (H22) study revealed that bortezomib via tailvein can obviously inhibit the growth of the tumor by inducing tumor cell’s apoptosis.Compared with control group: The tumor volume of experimental mice were significantlysmaller (P <0.05); the ratio of tumor size/mouse weight of experimental mice weresignificantly lower (P <0.05);In immunohistochemical assay: Compared with controlgroup: the expression of TNF@(apoptosis promoting protein) of experiment group wasobviously higher and the expression of apoptosis inhibiting protein (XIAP, SURVIVIN,STAT3) were significantly decreased (P <0.05).Part2Study of hepatoma cell lines transfected with plasmid containGFP and ISG15and following expressionObjective: Sieve the hepatocellular carcinoma cell lines with stable expression ofGFP(green fluorescence protein)and observe the transfection of ISG15contained plasmid and following expressionMethods: Transfect the GFP and ISG15plasmid into hepatoma cell lines,fluorescence microscopy and Western Blotting detection camera ISG15expression levels,and selected with G418with fluorescent and stable expression of GFP hepatoma cell lines.Results: After PcDNA3.1-GFP plasmid and PcDNA4.0-ISG15plasmid intohepatoma cells, fluorescence microscopy and Western Blotting Tips photographedsuccessful transfection and G418selection with the best drug fluorescence hepatoma cellline was maintained at a concentration of400ug/ml.After transfection the expression ofISG15was elevated significantly (P<0.05),but the selection for the hepatoma cell linesstablely express GFP(green fluorescence protein) by G418was end in fallure.Conclusions: Lipofectamine transfect system is suitable for plasmid transfection anddetection the expression of the corresponding pruducts in hepatic cancer cell,but due to thelow transfect effeciency and cell toxicity it’s not the best candidate for stable transfection use.