Study On The Molecular Mechanism Of Acute Inflammation In Suppressing MVD, Expression & Activity Of Gelatinase In Melanoma

ObjectiveTo investigate the effect of acute inflammation on melanoma growth through volume,microvessel density(MVD), expression and activity of matrix metalloproteinase (MMP)-2 and MMP-9.Contents1.The comparison of various acute inflammation-melanoma mice model.There are four types of acute inflammation-melanoma complex mice models:Simple wound cutting, repeated wound cutting, wound cutting plus staphylococcus aureus, wound cutting plus glacial acetic acid.To analyse advantages and disadvantages of each model.Chosing the most appropriate for next stage.2.The effect of acute inflammation on growth of tumor's volume. To build melanma-repeated wound cutting model in mice,invesgate tumor groth curve everday,then analyse tumor's volume of each group3.The effect of acute inflammation on growth of endothelium-dependent vascular. To build melanma-repeated wound cutting model in mice,all mice were killed 4 days after applying first wound cutting,then analyse MVD of each group4.The effect of acute inflammation on expression of MMP-2 and MMP-9.To build melanma-repeated wound cutting model in mice,all mice were killed 4 days after applying first wound cutting,then analyse expression and activity of MMP-2 and MMP-9 in each group.Methods1.B16F10 melanoma cells suspention was injiected into the left groin area of the mice at 0.2ml for each C57BL mouse. When the tumor were about 500mm3 four different wounds were built on the opposite side of bodies in the experiment groups.2. Invesgating wound healing, recording long diameter and short of every tumor, drawing tumor groth curve everday.3.To build melanoma-repeated cutting mice model according result of pre-experiment.4.4 days after applying first wound cutting, all mice were killed to get 40 cases tumor specimens,20 cases lung specimens.5.Tumor's volume was calculated accuording literature6.MVD were examined by Hematoxylin-Eosin stain and counted by Weidner method.7. Expression of MMP-2 and MMP-9 were examined by immunohistochemistry staining and analysed by Remmele-Stegner method.Activity of MMP-2 and MMP-9 were examined by gelatin zymography and analysed by Phoretix 1 system.Results1.The simple wound cutting has no effect on tumor growth, the repeated wound cutting, the wound cutting plus staphylococcus aureus, the wound cutting plus glacial acetic acid can suppress growth of tumor,and contribute equally to tumor development.2.The difference of the average tumor volume in the experiment group and control group was statistically different.3.MVD of the experiment was 16.560±1.325,MVD of the control was 23.240±2.289,the difference of the average tumor MVD in the experiment group and control group was statistically different4. The positive expression ratio of MMP-2 and MMP-9 in experimeent group was statistically different from the control group (p<0.05), The activity of MMP-2 and MMP-9 in experiment group was statistically different from the control group.Conclusion1.The simple wound have no effect on tumor growth;The repeated wound cutting, the wound cutting plus staphylococcus aureus, the wound cutting plus glacial acetic acid can decrease tumor development.Acute can suppress growth of tumor if it could sustain a utility level and last a period of time.2.The repeated wound cutting, the wound cutting plus staphylococcus aureus, the wound cutting plus glacial acetic acid have same influence on tumor growth, but they have different request on operation, management and feeding.The repeated wound cutting was applied for study on MVD,the expression and activity of MMP-2 and MMP-9.3.Acute is able to inhibit the expression and activity of MMP-2 and MMP-9, leading to decrease of endothelium-dependent vascular,which ultimately inhibits the growth of B16 melanoma.