Enhancement of Tumor Growth and Vascular Density by Transfection of Vascular Endothelial Cell Growth Factor Into MCF–7 Human Breast Carcinoma Cells (original) (raw)

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Affiliations of authors: Molecular Angiogenesis Group, Imperial Cancer Research Fund, Institute of Molecular Medicine. University of Oxford, John Radcliffe Hospital

England

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Nuffield Department of Pathology, University of Oxford, John Radcliffe Hospital

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Affiliations of authors: Molecular Angiogenesis Group, Imperial Cancer Research Fund, Institute of Molecular Medicine. University of Oxford, John Radcliffe Hospital

England

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Department of Pharmacology, University of Firenze

Italy

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Department of Gene Expression, G. F. Biotechnologische Forschung, Braunschweig, Federal Republic of Germany

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Affiliations of authors: Molecular Angiogenesis Group, Imperial Cancer Research Fund, Institute of Molecular Medicine. University of Oxford, John Radcliffe Hospital

England

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Correspondence to: Roy Bicknell, D.Phil., Molecular Angiogenesis Group, Imperial Cancer Research Fund, Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford, OX3 9DU, England.

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Published:

01 February 1995

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Hua-Tang Zhang, Paul Craft, Prudence A. E. Scott, Marina Ziche, Herbert A. Weich, Adrian L. Harris, Roy Bicknell, Enhancement of Tumor Growth and Vascular Density by Transfection of Vascular Endothelial Cell Growth Factor Into MCF–7 Human Breast Carcinoma Cells, JNCI: Journal of the National Cancer Institute, Volume 87, Issue 3, February 1995, Pages 213–219, https://doi.org/10.1093/jnci/87.3.213
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Abstract

Background : Vascular endothelia growth factor (VEGF) is a secreted endothelial-specific growth factor that is angiogenic in vivo. it is commonly expressed in a range of carcinomas. Purpose : The study was designed to investigate the effect of constitutive expression of VEGF on tumor formation by estrogen-dependent human MCF–7 breast carcinoma cells. Methods : A full-length complementary DNA encoding the shortest isoform of VEGF (VEGF 121 was stably transfected into MCF–7 cells. transfected clones were screened for VEGF 121 messenger RNA (mRNA) expression by ribonuclease protection analysis and for secretion of VEGF 121 protein by Western blot analysis. secretion of biologically active VEGF 121 by transfectants was confirmed by 1) a competitive radioreceptor-binding assay, 2) stimulation of the growth of microvascular endothelial cells in vitro, and 3) potent angiogenic activity in the rabbit corneal assay. tumor models were then established by subcutaneously implanting wildtype of VEGF 121 -transfected MCF–7 cells, together with either mouse BALB/3T3 clone A31 fibroblasts or human MDA–435S breast carcinoma cells, into ovariectomized nude mice either with or without a separately implanted slow-release estrogen pellet. Tumor vascularity was quantitatively assessed by capillary vessel counting after staining with the pan-endothelia marker CD31. Results : Stable VEGF 121 -overexpressing MCF–7cells were isolated and designated V12 cells. when implanted into the rabbit cornea, V12 cells elicited a strong directional outgrowth of capillaries. The growth rate of V12 cells in vitro was indistinguishable from that of MCF–7 wild—type cells. V12 cells formed faster growting tumors than did wild-type cells (P←.01) and showed a heterogeneous distribution of vessels when compared with the homogeneous distribution seen in tumors formed form wild-type cells. VEGF 121 overexpression had no effect on hormone dependence or tamoxifen sensitivity of tumor formation by MCF–7 cells in mice. No macroscopic evidence for metastasis from subcutaneous implants was obtained conclusions : VEGF 121 expression by breast carcinoma cells confers a growth advantange in vivo but not in vitro. Tumors formed by V12 transfectants were more vascular than those formed by wild-type MCF–7 cells, and we surmise that the growth advantage arises from increased tumor vascularization induced by VEGF 121.Implications : Tumor formation by V12 cells could provide a useful model for the assessment of anti-angiogenic drugs. [J Natl Cancer Inst 87: 213–219, 1995]

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