Summary
Vascular endothelial growth factor (VEGF) is recognized to play a predominant role in breast cancer prognosis. The action of VEGF is mediated by two high-affinity receptors with ligand-stimulated tyrosine kinase activity: VEGFR-1/flt-1 and VEGFR-2/flk-1, which are expressed mainly in vascular endothelial cells. To the best of our knowledge, no previous studies on the expression of these receptors in breast cancer cells has been made. We have established a new animal model for breast cancer, using a combination of 17β-oestradiol and testosterone as ‘carcinogens’. Taking advantage of the animal model, we have demonstrated that mammary cancer cells expressed not only high levels of VEGF but also, surprisingly, its receptors (flt-1 and flk-1) in mammary cancer cells. Intense reactivities to VEGF, flt-1 and flk-1 were observed in mammary cancer cells, especially in invasive mammary carcinoma. Western blot analysis confirmed the increase in flk-1 and flt-1 proteins in induced mammary cancers. Based on these observations, we hypothesize that in mammary cancer, VEGF regulates, in addition to endothelial proliferation and angiogenesis, also growth of cancer cells by an autocrine mechanism mediated through its receptors. To further verify this hypothesis, we investigated the correlation between cellular proliferation and the expression of VEGF, flt-1 and flk-1. Using double-labelling immunocytochemistry, we have shown a correlation between high VEGF activity and Ki-67 expression. The Ki-67 indices in the areas of strong and weak VEGF reactivities were 58.3% and 3.7% respectively. Similarly, there was also a correlation of strong flk-1 and Ki-67 reactivity. The Ki-67 indices for areas of strong and weak flk-1 reactivities were 53.9% and 3.1% respectively. On the other hand, there was a reverse correlation between flt-1 and Ki-67 activities. These results indicate that overexpression of VEGF and flk-1 is correlated with high Ki-67 index. The data, therefore, suggest that VEGF may act as an autocrine growth factor for mammary cancer cells in vivo and this autocrine regulatory role may be mediated through flk-1. The present study is the first report showing that VEGF may act as a growth stimulator for mammary cancer cells.
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Dellas, A, Schultheiss, E, Oberholzer, M, Torhorst, J & Gudat, F (1996). Analysis of proliferative activity using Ki-67 on cervical precancerous lesions and the relationship to p53 expression. Anticancer Res 16: 3403–3408.
Dvorak, H, Brown, L, Detmar, M & Dvorak, A (1995). Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. Am J Pathol 146: 1029–1039.
Enomoto, T, Okamoto, T & Sato, JD (1994). Vascular endothelial growth factor induces the disorganization of actin stress fibers accompanied by protein tyrosine phosphorylation and morphological change in Balb/C3T3 cells. Biochem Biophys Res Commun 202: 1716–1723.
Ferrara, N (1995). The role of vascular endothelial growth factor in pathological angiogenesis. Breast Cancer Res Treat 36: 127–137.
Ferrara, N, Houck, KA, Jakeman, LB & Leung, DW (1991). The vascular endothelial growth factor family of polypeptides. J Cell Biochem 47: 211–218.
Fong, GH, Rossant, J, Gertsenstein, M & Breitman, ML (1995). Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium. Nature 376: 66–70.
Fox, SB & Harris, AL (1997). Markers of tumor angiogenesis: clinical applications in prognosis and anti-angiogenic therapy. Invest New Drugs 15: 15–28.
Gasparini, G, Toi, M, Gion, M, Verderio, P, Dittadi, R, Hanatani, M, Matsubara, I, Vinante, O, Bonoldi, E, Boracchi, P, Gatti, C, Suzuki, H & Tominaga, T (1997). Prognostic significance of vascular endothelial growth factor protein in node-negative breast carcinoma. J Natl Cancer Inst 89: 139–147.
Gerdes, J, Lelle, RJ & Pickartz, H (1986). Growth fractions in breast cancer determined in situ with the monoclonal antibody Ki-67. J Clin Pathol 39: 977–980.
Gitay-Goren, H, Halaban, R & Neufeld, G (1993). Human melanoma cells but not normal melanocytes express vascular endothelial growth factor receptors. Biochem Biophys Res Commun 190: 702–708.
Hanahan, D (1997). Signaling vascular morphogenesis and maintenance. Science 277: 48–50.
Hanahan, D & Folkman, J (1996). Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell 86: 353–364.
Hendrix, MJC, Muschel, RJ & Padarathsingh, M (1997). Recent advances in breast cancer research: from genes to management. Am J Pathol 151: 883–888.
Martin, L, Green, B, Renshaw, C, Lowe, D, Rudland, P, Leinster, SJ & Winstanley, L (1997). Examining the technique of angiogenesis assessment in invasive breast cancer. Br J Cancer 76: 1046–1054.
Masood, R, Cai, J, Zheng, T, Smith, DL, Naidu, Y & Gill, PS (1997). Vascular endothelial growth factor/vascular permeability factor in an autocrine growth factor for AIDS-Kaposi sarcoma. Proc Natl Acad Sci USA 94: 979–984.
Millauer, B, Wizigmann-Voos, S, Schnurch, H, Martinez, R, Moller, NPH, Risau, W & Ullrich, A (1993). High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis. Cell 72: 835–846.
Mitola, S, Sozzani, S, Luini, W, Primo, L, Borsatti, A, Weich, H & Bussolino, F (1997). Tat-human immunodeficiency virus-1 induces human monocyte chemotaxis by activation of vascular endothelial growth factor receptor-1. Blood 90: 1365–1372.
Parker, SL, Tong, T, Bolden, S & Wingo, PA (1997). Cancer statistics 1997. CA Cancer J Clin 47: 5–27.
Plate, KH, Breier, G, Millauer, B, Ullrich, A & Risau, W (1993). Up-regulation of vascular endothelial growth factor and its cognate receptors in a rat glioma model of tumor angiogenesis. Cancer Res 53: 5822–5827.
Plate, KH, Breier, G, Weich, HA, Mennel, HD & Risau, W (1994). Vascular endothelial growth factor and glioma angiogenesis: coordinate induction of VEGF receptors, distribution of VEGF protein and possible in vivo regulatory mechanisms. Int J Cancer 59: 520–529.
Ref, M, Lejeune, S, Scott, PA, Fox, S, Smith, K, Leek, R, Moghaddam, A, Whitehouse, R, Bicknell, R & Harris, AL (1997). Expression of the angiogenic factors vascular endothelial cell growth factor, acidic and basic fibroblast growth factor, tumor growth factor beta-1, platelet-derived endothelial cell growth factor, placenta growth factor, and pleiotrophin in human primary breast cancer and its relation to angiogenesis. Cancer Res 57: 963–969.
Rose, DSC, Maddox, PH & Brown, DC (1994). Which proliferation markers for routine immunohistology? A comparison of five antibodies. J Clin Pathol 47: 1010–1014.
Ross, W & Hall, PA (1995). Ki-67: from antibody to molecule to understanding. J Clin Pathol:Clin Mol Pathol 218: M113–M117.
Russo, J, Russo, IH, Rogers, AE, Van Zwietan, MJ & Gusterson, B (1990). Tumors of the mammary gland. IARC Scientific Publications 99: 47–78.
Seetharam, L, Gotoh, N, Maru, Y, Neufeld, G, Yamaguchi, S & Shibuya, M (1995). A unique signal transduction from FLT tyrosine kinase, a receptor for vascular endothelial growth factor VEGF. Oncogene 10: 135–147.
Senger, DR, Galli, SJ, Dvorak, AM, Perruzzi, CA, Harvey, VS & Dvorak, HF (1983). Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. Science (Washington DC) 219: 983–985.
Senger, DR, Van De Water, L, Brown, L, Nagy, J, Yeo, K-T, Yeo, T-K, Berse, B, Jackman, R, Dvorak, A & Dvorak, H (1993). Vascular permeability factor (VPF, VEGF) in tumor biology. Cancer Metastasis Rev 12: 303–324.
Shibuya, M (1995). Role of VEGF-Flt receptor system in noral and tumor angiogenesis. Adv Cancer Res 67: 281–316.
Shim, JW, Koh, YC, Ahn, HK, Park, YE, Hwang, DY & Chi, JG (1996). Expression of bFGF and VEGF in brain astrocytoma. Journal of Korean Medical Science 11: 149–157.
Takahashi, T & Shibuya, M (1997). The 230 kDa mature form of KDR/Flk-1 (VEGF receptor-2) activates the PLC-gamma pathway and partially induces mitotic signals in NIH3T3 fibroblasts. Oncogene 14: 2079–2089.
Takahashi, Y, Kitadai, Y, Bucana, CD, Cleary, KR & Ellis, LM (1995). Expression of vascular endothelial growth factor and its receptor, KDR, correlates with vascularity, metastasis, and proliferation of human colon cancer. Cancer Res 55: 3964–3968.
Tio, M, Hoshina, S, Takayanagi, T & Tominaga, T (1994). Association of vascular endothelial growth factor expression with tumor angiogenesis and with early relapse in primary breast cancer. Jpn J Cancer Res 85: 1045–1049.
Waltenberger, J, Claesso-Welsh, L, Siegbahn, A, Shibuya, M & Heldin, CH (1994). Different signal transduction properties of KDR and FLT1, two receptors for vascular endothelial growth factor. J Biol Chem 269: 26988–26995.
Wintzer, H-O, Schulte-Monting, J, Hellerich, U & Von Kleist, S (1991). Ki-67 immunostaining in human breast tumors and its relationship to prognosis. Cancer 67: 421–428.
Wong, YC, Xie, B & Tsao, GSW (1999). Induction of breast cancer in Noble rats with a combination of estrogen and testosterone. In:Hormonal Carcinogenesis III, Li JJ Daling J Li SASpringer-Verlag: New York
Xie, B, Tsao, SW & Wong, YC (1999a). Induction of high incidence of mammary tumor in female Noble rats with a combination of 17β-oestradiol and testosterone. Carcinogenesis 20, (in press)
Xie, B, Tsao, SW & Wong, YC (1999b). Sex hormone-induced mammary carcinogenesis in the female Noble rats: the role of androgens. Carcinogenesis 20: 1069–1078.
Xie, B, Tsao, SW & Wong, YC (1999c). Sex hormone-induced mammary carcinogenesis in the female Noble rats: expression of TGF-β1 and its receptors, TGF-α, and EGF-R in mammary carcinogenesis. Breast Cancer Res Treat, 1597–1606.
Xie, B, Tsao, SW & Wong, YC (1999d). Isolation and purification of breast carcinoma cell lines from a Noble rat. In:Hormonal Carcinogenesis III, Li JJ Daling J Li SASpringer-Verlag: New York
Yamane, A, Seetharam, L, Yamaguchi, S, Gotoh, N, Takahashi, T, Neufeld, G & Shibuya, M (1994). A new communication system between hepatocytes and sinusoidal endothelial cells in liver through vascular endothelial growth factor and Flt tyrosine kinase receptor family (Flt-1 and KDR/Flk-1). Oncogene 9: 2683–2690.
Yang, X & Cepko, CL (1996). Flk-1, a receptor for vascular endothelial growth factor (VEGF) is expressed by retinal progenitor cells. J Neurosci 16: 6089–6099.
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Xie, B., Tam, N., Tsao, S. et al. Co-expression of vascular endothelial growth factor (VEGF) and its receptors (flk-1 and flt-1) in hormone-induced mammary cancer in the Noble rat. Br J Cancer 81, 1335–1343 (1999). https://doi.org/10.1038/sj.bjc.6692206
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bjc.6692206
Keywords
This article is cited by
-
Modulation of VEGF/Flk-1 receptor expression in the rat pituitary GH3 cell line by growth factors
Pituitary (2006)
-
Effects of oestradiol and tamoxifen on VEGF, soluble VEGFR-1, and VEGFR-2 in breast cancer and endothelial cells
British Journal of Cancer (2005)
-
Inhibition of VEGF receptors significantly impairs mammary cancer growth in C3(1)/Tag transgenic mice through antiangiogenic and non-antiangiogenic mechanisms
Oncogene (2005)
-
Sex steroid regulation of angiogenesis in breast tissue
Angiogenesis (2005)
-
Tumor-specific VEGF-A and VEGFR2 in postmenopausal breast cancer patients with long-term follow-up. Implication of a link between VEGF pathway and tamoxifen response
Breast Cancer Research and Treatment (2005)