Experimental Study About Inhibition Of The Invasion Effect Of Anti-EMMPRINmAb On TSCC

Objective: To explore the mechanism of blocking the tumor erodes and search for a new way to treat the oral squamous cell carcinoma.we choose Anti-EMMPRIN monoclonal antibody plus matrix metalloproteinases inhibitors Doxycycline on the proliferation of Tca8113cells. We also establish TSCC model through nude mouse to explore the effect of the drugs.Methods:1. MTT (Methyl thiazolyl tetrazolium) assay was applied to evaluating the inhibitory effects of Anti-EMMPRIN monoclonal antibody and Doxycycline on the Tca8113cells during different time.2. BALB/c nude mice were used to establish tongue squamous cell carcinoma model by subcutaneously injection of1×106Tca8113cells.Two weeks later, thirty-two tumor-bearing mice were randomly divided into four groups, the control group,the Anti-EMMPRIN monoclonal antibody group, the Doxycycline group and the combination group,the number of mice in each group was8. The control group was given saline injection. The Anti-EMMPRIN monoclonal antibody group was given Anti-EMMPRIN monoclonal antibody10mg/kg/d. The Doxycycline group was given20mg/kg/3d.The combination group was given Anti-EMMPRIN monoclonal antibody and Doxycycline. The tumor volumes and weights were observed and measured every three days. Calculating tumor inhibition rate.3. Detect the protein expression of CD147,MMP-2and MMP-9in the organization tumor by immunohistochemical stains. CD147and MMP-2positive signal located in the cell membrane and cell plasma, MMP-9positive signal located in the cytoplasm and cell membrane, into tan particles. Randomly selected four at high magnification the field of vision, count the positive cells, and the results are as follows:<1%(0),1%-10%(1),10%-50%(2),>50%(3), the before two are considered to be negative,the follow two are considered to be positive.Results:1. The result of cell proliferation test:The combination group could inhibit the proliferation of Tca8113in vitro, which is stronger than Anti-EMMPRIN monoclonal antibody group and Doxycycline group.2. The tumor weights and inhibit rates:The tumor weight of four groups were respectively1.12±0.31g,0.86+0.11g,0.74±0.19g and0.55±0.17g. There were significant differences among control group and other groups (P<0.01). The combination group and the singal group were different(P<0.05). The volume of control group is significant different to before, the other three groups are the opposite.There is significant differences among control group and experiment groups (P<0.01).The combination group changes smaller than Doxycycline group and Anti-EMMPRIN monoclonal antibody group(P<0.05).There is no difference among The Anti-EMMPRINmAb and the Doxycycline (P>0.05).The tumour inhibition rates are37.58%,34.28%and40.35%.3. The protein expression of CD147and MMP-2, MMP-9:the control group:CD147protein positive expression is100%, MMP-2protein positive expression is100%, MMP-9protein positive expression is100%. Anti-EMMPRIN monoclonal antibody group:CD147protein positive expression is25%, MMP-2protein positive expression is12.5%, MMP-9protein positive expression is12.5%. Doxycycline group:CD147protein positive expression is50%, MMP-2protein positive expression is50%, MMP-9protein positive expression is50%. The combination group:CD147protein positive expression is12.5%, MMP-2protein positive expression is12.5%, MMP-9protein positive expression is12.5%.