Biological Properties And Recombinant Human Vascular Endostatin Therapy Of Hemangioma: Study On Hemangioma Endothelial Cells In Vitro Culture

Objective To consummate the in vitro culture method of hemangioma endothelial cells and to observe the growth regulation and the biological character of hemangioma vascular endothelial cells. To study the mechanism of the hemangioma treatment by using recombinant human vascular endostatin.Methods The hemangioma vascular endothelial cells which came from the sterile hemangioma were cultured by tissue culture method. The expression of the factorâ…§related antigen which only exists in the endothelial cells to identify the hemangioma vascular endothelial cell (VEC) were observed by the Immunohistochemistry technique. After Recombinant human endostatin(YH-16)was used in hemangioma endothelial cells,the morphological changes were observed under the light microscope at different time points of 24h,48h,72h.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetra zolium bromide (MTT) was used to examine the inhibitory action of YH-16 on hemangioma endothelial cells proliferation. The flow cytometry technique was used to examine the effect of YH-16 on hemangioma endothelial cell apoptosis in the earlier period.Results The separated cells, as epithelium form, can be observed by the phase contrast microscope. The cells emigrated from the tissue to the postcultural after four days. The high-speed growth was found after three weeks .The expression factorâ…§associated antigen was positive in the isolated cells. AfterYH-16 was used in hemangioma endothelial cells, the disseminated necrosis was discovered under the light microscope. MTT assay showed that the absorbance values decreased with the increase of drug concentration (P<0.05).Compared with the lowest concentration group(15.625ug/ml),the same dose- effect relationship was found in higher groups(P<0.05).The inhibition rate increased with the increase of the drug concentration and time. Flow cytometry showed that the apoptosis rate of hemangioma endothelial cells in the early period gradually increased with the increase of drug concentration(P<0.05).Compared with the lowest concentration group(15.625ug/ml),the same dose-effect relationship was found in higher groups(P<0.05).Conclusion Tissue culture method was a simple way to make the hemangioma endothelial cell alive.YH-16 can inhibit hemangioma endothelial cell proliferation, as well as promoting apoptosis.