The Mechanisms Of Heparanase Inducd Capillary Capture In Hepatocellular Carcinoma

Object: To expore the mechanism of heparanase(HPSE)-induced cell adhesion between hepatocellular carcinoma(HCC)cells and capillary endothelial cells.In order to discover the key heparan sulfate proteoglycan(HSPG)involved in the adhesion procession and its distribution in the cell.Method :1)The HPSE gene interference sequence of si HPSE-3158 was selected to construct recombinant vector,then transfected into the Hep G-2 cells with lipofectin transfection to inhibit HPSE gene expression.Meanwhile,negative control group and blank control group were set for comparison.After the adhesion between Hep G-2cells and endothelial cells Human Umbilical Vein Endothelisl Cells(HUVEC)in different groups in 6-well plate,the expression of syndeccan-1 ? syndeccan-3 of HSPGs were detected by western-blot.2)HSPG subtypes were selected according to its expression change,and the cellular localization were observed by immunocytochemistry.Result:1)The result of Western-Blot:After the adhesion between Hep G-2 cells and HUVEC cells,the expression of Syndecan-1 showed in relative grey value was1.885±0.113?1.870±0.087?1.837±0.116 in the transfected group ?the negative control group and blank control group which showed that the Syndecan-1 expression has no significant difference in those groups(P>0.05).The Syndecan-3 expression showed in relative grey value was 1.452±0.093?1.079±0.134?1.019±0.105,thus we can find that the Syndecan-3 expression was higher in the transfected group compared with the other two groups(P>0.05)but showed no difference between negative control group and blank control group(P<0.05).2)Immunohistochemical result :Syndecan-3 was stained in the HUVEC cytoplasm and cell membrane.Conclusion : In the process of liver cancer cells and vascular endothelial cells adhesion,HPSE has a certain influence on HSPG expression.Syndecan-3 might play an important role in the HPSE induced adhesion between HCC cells and capillary endothelial cells,and located mainly in the HUVEC cytoplasm and cell membrane.