To establish the immortalized human umbilical vein endothelial cell lines and investigate the effect of immortalization on biological characteristics of proliferation, apoptosis and tumorigenesis of the cells.

Plasmid containing simian virus 40 large T antigen (SV40LT) gene or human telomerase reverse transcriptase (hTERT) gene was amplified and extracted; the target genes marked by puromycin resistance were packed by human embryo kidney cells (293T) to get the retroviral vectors and used to infect human umbilical vein endothelial cells (HUVECs); then the infected cells were screened by culture medium containing 2 μg/ml puromycin for two weeks; and polymerase chain reaction (PCR) was used to identify whether the target genes were successfully transferred into HUVECs; cell counting method, flow cytometry and tumorigenesis experiments in nude mice were used to detect the biological characteristics of proliferation, apoptosis and malignant transformation of immortalized cells respectively.

The human umbilical vein endothelial cells were successfully immortalized by retrovirus-mediated SV40LT and hTERT; On the 5th to the 7th day, there were significant differences in cells number between the SV40LT immortalized group and HUVECs group (P<0.05) and between the hTERT immortalized group and HUVECs group (P<0.05). The apoptosis rate of HUVECs group was (11.2±0.9)%, the apoptosis rate of HUVECs immortalized by SV40LT was (3.4±0.4)%, the apoptosis rate of HUVECs immortalized by hTERT was (2.8±0.4)%. The apoptotic rates of the two immortalized group were lower than that of HUVECs group (bothP<0.01). These immortalized HUVECs had no tumorigenicity.

Retrovirus-mediated SV40LT and hTERT can successfully immortalize HUVECs; the proliferative capacity of immortalized HUVECs is greatly enhanced; the apoptosis rate is decreased, and no malignant transformation occurs.

Simian virus 40 large T antigen; Human telomerase reverse transcriptase; Retroviral vector; Human umbilical vein endothelial cell; Immortalization