| Objective: To construct the RNAi interference hepatoma cell line He PG-2 were observed by a variety of different effects on the expression of heparanase expression in hepatocellular carcinoma cell adhesion molecules on vascular endothelial cells,improve and enrich the molecular study of heparanase in invasion and metastasis of hepatocellular carcinoma.Methods:(1)The hepatocellular carcinoma cell line He PG-2 as the experimental cell research,the inhibition of heparanase gene transfection on He PG-2 cells,HPSE transfected cells were detected by q RT-PCR and Western-blot methods of gene expression level;(2)by the above-mentioned method to choose the corresponding He PG-2 cells were divided into transfection group(RNAi interference of He PG-2 cells),negative control group(transfected negative He PG-2 cells)and control group(non transfected He PG-2 cells)cells in three groups of cell adhesion experiments,analyzing the effect of various experimental group of cell adhesion molecules expression by q RTPCR and Western-blot method.Results:(1)Using q RT-PCR and Western-blot experimental analysis method selection,successfully obtained the optimal interference of He PG-2 cells;(2)the q RT-PCR and Western-blot results: the He PG-2 cells transfected with enhanced expression of adhesion molecules on vascular endothelial cells;RNAi interference of He PG-2 cells significantly decreased the expression of adhesion molecules on vascular endothelial cells;transfection the negative expression of He PG-2 cells compared with the blank control group decreased adhesion molecules on vascular endothelial cells,cells in He PG-2 group but higher than that of the RNAi interference.Conclusion: The q RT-PCR and Western-blot experimental method that heparanase played on hepatoma cells and adhesion in endothelial cells during the course of the important role that mediate cell adhesion of hepatocellular carcinoma cells and vascular endothelial cells. |
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