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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 volume 21, pages 4200–4211 (2002)Cite this article

Abstract

We have modelled multiple stages of malignant transformation of human endothelial cells (ECs) by overexpressing the catalytic subunit of human telomerase (hTERT), together with SV40 T antigen (SV40T) and oncogenic N-ras. Transfection with hTERT alone, led to the immortalization of two out of three cultures of bone marrow-derived ECs (BMECs). One hTERT transduced BMEC culture underwent a long proliferative lag before resuming proliferation. BMECs transfected with hTERT alone were functionally and phenotypically normal. BMECs transfected with SV40T (BMSVTs) had an extended lifespan, but eventually succumbed to crisis. BMSVTs exhibited a partially transformed phenotype, demonstrating growth factor independence, altered antigen expression and forming tiny, infrequent colonies in vitro. Transduction of BMSVTs with hTERT resulted in immortalization of 4 out of 4 cultures. BMSVTs immortalized with hTERT formed large colonies in vitro and small transient tumours in vivo. BMECs co-expressing SV40T, hTERT and N-ras exhibited an overtly transformed phenotype; forming very large colonies with an altered morphology and generating rapidly growing tumours in vivo. These investigations demonstrate transformation of human ECs to an overtly malignant phenotype. This model will be useful for understanding mechanisms underlying vascular and angiogenic neoplasias, as well as for testing drugs designed to curtail aberrant EC growth.

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Acknowledgements

This work was supported by NCI grant CA59350, NHLBI grant HL 61401 and the Gar Reichman Fund of Cancer Research Institute. We thank Ms Dianna Ngok, Mr Jason Anselmo, Mr Sheik Baksh and Mr Harry Satterwaite for technical assistance, Mr George Nam for preparation of ECGM and EC isolation. We thank Geron Corporation for the hTERT cDNA and Imclone Systems Incorporated for antibodies to VEGF receptors.

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  1. Karen L MacKenzie

    Present address: Children's Cancer Institute Australia for Medical Research, High St., P.O. Box 81 Randwick, NSW, 2031, Australia

Authors and Affiliations

  1. James Ewing Laboratory of Developmental Haematopoiesis and Department of Human Genetics, Sloan-Kettering Cancer Institute, New York, NY, USA

    Karen L MacKenzie, Sonia Franco, Chad May, Michel Sadelain & Malcolm A S Moore

  2. Cornell University Medical College, New York, NY, USA

    Afzal J Naiyer & Shahin Rafii

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Correspondence to Karen L MacKenzie.

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MacKenzie, K., Franco, S., Naiyer, A. et al. 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, 4200–4211 (2002). https://doi.org/10.1038/sj.onc.1205425

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