Isolation And Identification Of Genes Associated With Tumor Endothelial Cells

The growth and metastasis of tumor are dependent upon tumor angiogenesis. Inhibition of tumor angiogenesis is a new anticancer strategy. Tumor endothelial cells play an important role in tumor angiogenesis. They are the targets of many angiogenic factors. Drugs targeting the endothelial cells could kill endothelial cells and destroy blood vessels of tumor.s. We here isolated and identified some tumor endothelial cell- related genes first, and then studied the preliminary function of two unknown genes.Isolation of Genes Associated with Tumor Endothelial CellsHuman umbilical vein endothelial cells(HUVECs) were isolated, cultured and treated with tumor tissues homogenate, VEGF (Vascular Endothalial Growth Factor)and bFGF (basic Fibroblast Growth Factor), HUVECs were stimulated as activated HUVECs. mRNA was isolated from activated HUVECs, A primary cDNA library consisting of 4 × 10⁶ recombinants was constructed. After the suppression subtractive hybridization (SSH) was performed between normal HUVECs and activated HUVECs, the different gene fragments of activated HUVECs were obtained and labeled with ³²P by random primers method to make the probes. The cDNA library was screened with the probes and 177 positive clones were obtained, from which 70 clones were selected randomly and sequenced. Bioinformational analysis of DNA sequences indicated that 70 positive clones represent 35 EST ,which can be classified as the following: A) 11 of genes associated with tumor...