Establishment and Identif ication of Tumor Vascular Endothelial Cell Model

Objective 　To establish the cell model of vascular endothelial cell in tumors and explore its biological features. Methods 　The normal human umbilical vein endothelial cell ECV304 was cultured with the supernatant of human ovarian cancer cell SKOV3 for three and half months, and ECV304-SKOV3 cells with stable growth was acquired. Then the changes of their biological characteristics were determined by light microscopy, electron microscopy, karyotype analysis, MTT assay, and flow cytometry. The expression of proliferating cell nuclear antigen ( PCNA), bcl-2, tenascin ( TN), matrix metalloproteinase 9 (MMP-9), and telomerase activity was detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR) respectively. The ability to synthesize proteins was determined. Results 　Abnormal nuclei and compartments with microvilli were observed in ECV304-SKOV3 cells and their chromatosome modes increased slightly. As compared with parental cells, the population doubling time of ECV304-SKOV3 cells shortened, mitotic indices increased, the expression of PCNA, bcl-2 and MMP-9 enhanced (t ≥2. 4671, P<0. 01), while TN decreased ( t=2. 2473, P <0. 05), the cell number in S-phase increased, while in G0 >>/ G1>>-phase decreased significantly (χ2 = 923. 313, P < 0. 01) . ECV304-SK-OV3 cells were more sensitive than ECV304 to anticancer drugs with inhibition effect on S-phase or S-and M-phase, in accordance with SKOV3 cells ( F≥24. 14, P<0. 01) . The ability to synthesize proteins and telomerase activity enhanced ( t=-12. 2465, P<0. 001 ; t =-25. 2990, P<0. 001) . Conclusion 　The result s suggest that ECV3042SKOV3 cells have some biological characteristics of vascular endothelial cells in tumors, including proliferating more quickly, surviving longer and more functional, expressing some angiogeneic factors st ronger, and can act as a cell model for developing drugs targeting the tumor vascular endothelial cells in the future.