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Received: 2004-09-06

Revision Accepted: 2005-01-27

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Journal of Zhejiang University SCIENCE B 2005 Vol.6 No.7 P.631-636


Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen

Author(s):  BIAN Chang, ZHAO Kui, TONG Guo-xin, ZHU Yong-liang, CHEN Peng

Affiliation(s):  Department of Cardiology, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China; more

Corresponding email(s):   skyoutwin@yahoo.com.cn

Key Words:  Endothelial cell, Telomerase activity, Immortalization

BIAN Chang, ZHAO Kui, TONG Guo-xin, ZHU Yong-liang, CHEN Peng. Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen[J]. Journal of Zhejiang University Science B, 2005, 6(7): 631-636.

@article{title="Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen",
author="BIAN Chang, ZHAO Kui, TONG Guo-xin, ZHU Yong-liang, CHEN Peng",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen
%A BIAN Chang
%A TONG Guo-xin
%A ZHU Yong-liang
%A CHEN Peng
%J Journal of Zhejiang University SCIENCE B
%V 6
%N 7
%P 631-636
%@ 1673-1581
%D 2005
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2005.B0631

T1 - Immortalization of human umbilical vein endothelial cells with telomerase reverse transcriptase and simian virus 40 large T antigen
A1 - BIAN Chang
A1 - ZHAO Kui
A1 - TONG Guo-xin
A1 - ZHU Yong-liang
A1 - CHEN Peng
J0 - Journal of Zhejiang University Science B
VL - 6
IS - 7
SP - 631
EP - 636
%@ 1673-1581
Y1 - 2005
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2005.B0631

Objective: To establish normally conditionally-immortalized human umbilical vein endothelial cells (HUVECs) by ectopic expression of the human telomerase catalytic enzyme (hTERT) and simian virus 40 large T (SV40 LT) antigen. Methods: Primary HUVECs were transfected with recombinant retrovirus containing hTERT or SV40 LT respectively. Subsequently drug resistant cell clones were screened and expanded for further studies. endothelial cell biomarkers were confirmed by examination. Results: The morphological phenotype of the transfected cells was similar to the non-transfected cells. Von Willebrand factor, hTERT and SV40 LT could be detected in transfected HUVECs. Moreover, higher telomerase activity in transfected cells was maintained for over 50 population doublings compared with only low level of endogenous telomerase transiently at early population doublings in primary HUVECs. When exposed to TNF-α (tumor necrosis factor-α), the expression of E-selectin in transfected cells was significantly up-regulated, but no alteration of endothelial lipase was found. Conclusion: Ectopic coexpression of hTERT and SV40 LT can effectively immortalize HUVECs without tumorigenicity in vitro. Immortalized HUVECs may be an ideal target of further molecular function studies.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1] Bodnar, A.G., Ouellette, M., Frolkis, M., Holt, S.E., Chiu, C.P., Morin, G.B., Harley, C.B., Shay, J.W., Lichtsteiner, S., Wright, W.E., 1998. Extension of life-span by introduction of telomerase into normal human cells. Science, 279(5349):349-352.

[2] Bruneel, A., Labas, V., Mailloux, A., Sharma, S., Vinh, J., Vaubourdolle, M., Baudin, B., 2003. Proteomic study of human umbilical vein endothelial cells in culture. Proteomics, 3(5):714-723.

[3] Burnham, E.L., Moss, M., Harris, F., Brown, L.A., 2004. Elevated plasma and lung endothelial selectin levels in patients with acute respiratory distress syndrome and a history of chronic alcohol abuse. Crit Care Med, 32(3):675-679.

[4] Charreau, B., Coupel, S., Goret, F., Pourcel, C., Soulillou, J.P., 2000. Association of glucocorticoids and cyclosporin A or rapamycin prevents E-selectin and IL-8 expression during LPS- and TNF alpha-mediated endothelial cell activation. Transplantation, 69(5):945-953.

[5] Che, W., Lerner-Marmarosh, N., Huang, Q., Osawa, M., Ohta, S., Yoshizumi, M., Glassman, M., Lee, J.D., Yan, C., Berk, B.C., et al., 2002. Insulin-like growth factor-1 enhances inflammatory responses in endothelial cells: Role of Gab1 and MEKK3 in TNF-(-induced c-Jun and NF-kappa (B) activation and adhesion molecule expression. Circ Res, 90(11):1222-1230.

[6] Darimont, C., Avanti, O., Tromvoukis, Y., Vautravers-Leone, P., Kurihara, N., Roodman, G.D., Colgin, L.M., Tullberg-Reinert, H., Pfeifer, A.M., Offord, E.A., et al., 2002. SV40 T antigen and telomerase are required to obtain immortalized human adult bone cells without loss of the differentiated phenotype. Cell Growth Differ, 13(2):59-67.

[7] Davies, B.R., Steele, I.A., Edmondson, R.J., Zwolinski, S.A., Saretzki, G., von Zglinicki, T., O’Hare, M.J., 2003. Immortalisation of human ovarian surface epithelium with telomerase and temperature-sensitive SV40 large T antigen. Exp Cell Res, 288(2):390-402.

[8] Greenwood, M.J., Lansdorp, P.M., 2003. Telomeres, telomerase, and hematopoietic stem cell biology. Arch Med Res, 34(6):489-495.

[9] Jaye, M., Lynch, K.J., Krawiec, J., Marchadier, D., Maugeais, C., Doan, K., South, V., Amin, D., Perrone, M., Rader, D.J., 1999. A novel endothelial-derived lipase that modulates HDL metabolism. Nat Genet, 21(4):424-428.

[10] Jha, K.K., Banga, S., Palejwala, V., Ozer, H.L., 1998. SV40-mediated immortalization. Exp Cell Res, 245(1):1-7.

[11] Jin, W., Sun, G.S., Marchadier, D., Octtaviani, E., Glick, J.M., Rader, D.J., 2003. Endothelial cells secrete triglyceride lipase and phospholipase activities in response to cytokines as a result of endothelial lipase. Circ Res, 92(6):644-650.

[12] Kudo, Y., Hiraoka, M., Kitagawa, S., Miyauchi, M., Kakuo, S., Zhao, M., Ide, T., Takata, T., 2002. Establishment of human cementifying fibroma cell lines by transfection with temperature-sensitive simian virus-40 T-antigen gene and hTERT gene. Bone, 30(5):712-717.

[13] Kumazaki, T., Hiyama, K., Takahashi, T., Omatsu, H., Tanimoto, K., Noguchi, T., Hiyama, E., Mitsui, Y., Nishiyama, M., 2004. Differential gene expressions during immortalization of normal human fibroblasts and endothelial cells transfected with human telomerase reverse transcriptase gene. Int J Oncol, 24(6):1435-1442.

[14] Lundberg, A.S., Randell, S.H., Stewart, S.A., Elenbaas, B., Hartwell, K.A., Brooks, M.W., Fleming, M.D., Olsen, J.C., Miller, S.W., Weinberg, R.A., et al., 2002. Immortalization and transformation of primary human airway epithelial cells by gene transfer. Oncogene, 21(29):4577-4586.

[15] Macera-Bloch, L., Houghton, J., Lenahan, M., Jha, K.K., Ozer, H.L., 2002. Termination of lifespan of SV40-transformed human fibroblasts in crisis is due to apoptosis. J Cell Physiol, 190(3):332-344.

[16] MacKenzie, K.L., Franco, S., Naiyer, A.J., May, C., Sadelain, M., Rafii, S., Moore, M.A., 2002. Multiple stages of malignant transformation of human endothelial cells modelled by co-expression of telomerase reverse transcriptase, SV40 T antigen and oncogenic N-ras. Oncogene, 21(27):4200-4211.

[17] Noble, J.R., Zhong, Z.H., Neumann, A.A., Melki, J.R., Clark, S.J., Reddel, R.R., 2004. Alterations in the p16(INK4a) and p53 tumor suppressor genes of hTERT-immortalized human fibroblasts. Oncogene, 23(17):3116-3121.

[18] O’Hare, M.J., Bond, J., Clarke, C., Takeuchi, Y., Atherton, A.J., Berry, C., Moody, J., Silver, A.R., Davies, D.C., Alsop, A.E., et al., 2001. Conditional immortalization of freshly isolated human mammary fibroblasts and endothelial cells. Proc Natl Acad Sci, 98(2):646-651.

[19] Proctor, C.J., Kirkwood, T.B., 2003. Modelling cellular senescence as a result of telomere state. Aging Cell, 2(3):151-157.

[20] Terasaki, T., Kyo, S., Takakura, M., Maida, Y., Tsuchiya, H., Tomita, K., Inoue, M., 2004. Analysis of telomerase activity and telomere length in bone and soft tissue tumors. Oncol Rep, 11(6):1307-1311.

[21] Veitonmaki, N., Fuxe, J., Hultdin, M., Roos, G., Pettersson, R.F., Cao, Y., 2003. Immortalization of bovine capillary endothelial cells by hTERT alone involves inactivation of endogenous p16INK4A/pRb. FASEB J, 17(6):764-766.

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