Liping Yang, Yixin Zhang, Wenjing Zhao, Chen Xudong, Qian Hongyan and Shao Jingjing
Key Laboratory, Cancer Research Center Nantong, Nantong University, Jiangsu 226361, China
Tumor angiogenesis plays a crucial role in cancer growth, recurrence and metastasis. However, the studies on the blood vessel formation and regulation in tumor have been hampered by the lack of a steady source of tumor endothelial cell (TEC). In our pilot study, we freshly isolated CD31+ endothelial cells from the tissues of human hepatocellular carcinoma (HCC) and cultured them in ECM medium with several supplements, and found that TECs in primary culture proliferated to confluency in 15 days to 20 days and stopped growing when subcultured to passage 7. To establish a novel TEC line, the cultured TECs were transfected with SV40-LT. The cloned TECs containing SV40-LT have been subcultured to passage 53 to date. The novel cell line of tumor endothelial cell derived from human HCC, with a steady growing rate in ECM medium has been established. These transfected TECs exhibited similar endothelial characteristics as the primary TECs, i.e. that appeared the cobblestone and spindle shape, Weibel-Palade bodies in the cells, both CD31 and CD105 expression on the cell surface, chromosome number with 44. Furthermore, functional assays showed that the tube formation and migration also occurred in the transfected cells. These results suggest that a novel TEC cell line established here maintains endothelial characteristics and potencies. Therefore, our TEC cell line should be useful for studying of tumor angiogenesis and vascular targeting therapy.
Acknowledgment: This study is granted by the Natural Science Foundation of China (NSFC), Grant No. 81272378.
Keywords: Tumor angiogenesis, tumor endothelial cell, cell line, human hepatocellular carcinoma.