Transformation of rabbit vascular smooth muscle cells by transfection with the early region of SV40 DNA.

Abstract : Rabbit aortic smooth muscle cells (SMC) and Rb-1 cells, a continuous line of the same origin, were transformed by transfection with pSV3-neo DNA, a plasmid containing the SV40 early region linked to the neoR resistance gene. Transformed clones were selected in G418-containing medium at a rate of 10(-4) per cell. All transformed clones were immortalized and contained in the early passages two free recombined plasmids derived from pSV3-neo. At advanced passages pSV3-neo sequences were found integrated in the cellular genome. Transformed cells had an altered morphology and growth pattern that differed among clones. Some clones reached high density in low-serum medium. All the clones stained positively for the intranuclear T antigen. Some clones had distinct transcripts for the large T and small t antigens, while in others only larger or truncated transcripts were found. Alpha-actin filaments were visualized by immunofluorescent staining in all the clones, but Northern blot analysis revealed a significant reduction in transcripts for this actin. All the transformed clones accumulated, to a variable extent, cholesteryl esters after incubation with beta very low-density lipoprotein. Six of the eight transformed clones maintained a diploid chromosome number, but there was an increase in structural chromosome aberrations, predominantly dicentrics. Transfection of pSV3-neo into rabbit vascular SMCs is an efficient model for obtaining transformed clonal populations. These clones show some phenotypic changes that may be relevant to the study of atherogenesis.
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Contributor : Laëtitia Legoupil <>
Submitted on : Thursday, May 23, 2019 - 6:03:06 PM
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  • HAL Id : hal-02138474, version 1
  • PUBMED : 2154928


M Nachtigal, Alain Legrand, M Nagpal, S Nachtigal, P Greenspan. Transformation of rabbit vascular smooth muscle cells by transfection with the early region of SV40 DNA.. American Journal of Pathology, American Society for Investigative Pathology, 1990, 136 (2), pp.297-306. ⟨hal-02138474⟩



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