Screening And Functional Studies Of Genes Associated With Human Liver Cancer Vascular Endothelial Cells

Angiogenesis is a critical event in solid tumor growth, invasion and metastasis. Tumor endothelium is a mediator during the complex process of tumor angiogenesis. Identifying and studying the function of genes associated with tumor endothelium can not only help to clarify the mechanism of tumor angiogenesis but also offer candidate targets for antiangiogenic therapeutic strategies of human tumors.Because of its rich vascular network, human liver cancer may be a proper disease for anti-angiogenic therapy. Selective inhibiting and directly killing human liver cancer endothelial cells may provide a valid treatment option for patients who are not suited to conventional therapies. Human hepatocellular carcinoma vascular endothelial cells (HCVECs) were isolated and purified from a fresh hepatocellular carcinoma tissue sample. A cDNA expression library with the size of 2×10~6pfu was constructed with the mRNA extracted from HCVECs. Then the identified xenogeneic sera from immunized mice with HCVEC were employed to screen the library of HCVECs by the method of modified serological analyses of recombinant cDNA expression libraries (SEREX). Thirty-six positive clones were obtained from 6×10~5 independent clones by immunoscreening. Bioinformatics analysis of cDNA sequences indicated that 36 positive clones represented 18 different genes. Among them, 3 of these genes were new genes previously unreported, 9 were associated with tumor angiogenesis, proliferation and migration of endothelial cells and tubular structure formation, and the other 6 may...