Summary
Production of vascular endothelial growth factor (VEGF) and expression of its receptors Flt-1 and KDR was determined in primary cultures of separated epithelial and stromal-enriched cultures derived from ten primary human breast carcinomas. By enzyme-linked immunosorbent assay, epithelial cells produced a mean VEGF of 33 ± 7 pg ml–1 μg–1 RNA (range 11–70). Stromal cells produced similar levels, with a mean of 48 ± 11 pg ml–1 μg–1 RNA (range 7–92). This was significantly greater than the amount produced by similar cultures derived from normal breast tissue (epithelial mean 19 ± 5 pg ml–1 μg–1 RNA, range 9–34, P < 0.05 vs tumour epithelial culture; stromal mean 26 ± 8 pg ml–1 μg–1 RNA, range 3–56). Flt-1 and KDR receptors were analysed by semi-quantitative reverse transcription polymerase chain reaction. Flt-1 was expressed by four of six epithelial and five of six stromal cultures. When expressed by both cell types, Flt-1 appeared to be significantly more abundant on stromal cells compared with epithelial cultures. Only a single tumour, a lobular carcinoma, failed to express Flt-1 on either cell type. With KDR, the reverse was true with constitutive expression of this receptor by epithelial cultures and zero or reduced (3/6) expression by stromal cultures. Differences in the expression pattern of VEGF receptors may reflect a differential response to VEGF by specific cell types. Thus, production of VEGF and expression of VEGF receptors Flt-1 and KDR by breast cancer epithelial and stromal cells suggests that VEGF may fulfil not only an angiogenic role, but also play a fundamental role as an autocrine/paracrine regulator in breast cancer, thereby facilitating tumour proliferation and subsequent invasion.
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
Adams, E. F., Rafferty, B. & White, M. C. (1991). Interleukin-6 is secreted by breast fibroblasts and stimulates 17β-estradiol oxidoreductase activity of MCF-7 cells: possible paracrine regulation of breast 17β-estradiol levels. Int J Cancer 49: 118–121.
Boocock, C. A., Charnock-Jones, S., Sharkey, A. M., McLaren, J., Barker, P. J., Wright, K. A., Twentyman, P. R. & Smith, S. K. (1995). Expression of vascular endothelial growth factor and its receptors flt and KDR in ovarian carcinoma. J Natl Cancer Inst 87: 506–516.
Brown, L. F., Berse, B., Jackman, R. W., Tognazzi, K., Guidi, A. J., Dvorak, H. F., Senger, D. R., Connolly, J. L. & Schnitt, S. J. (1995). Expression of vascular permeability factor (vascular endothelial growth factor) and its receptors in breast cancer. Hum Pathol 26: 86–91.
Charnock-Jones, D. S., Sharkey, A. M., Boocock, C. A., Ahmed, A., Plevin, R., Ferrara, N. & Smith, S. K. (1994). Vascular endothelial growth factor receptor localisation and activation in human trophoblast and choriocarcinoma cells. Biol Reprod 51: 524–530.
Cohen, T., Nahari, D., Cerem, L. W., Neufeld, G. & Levi, B-Z (1996). Interleukin-6 induces the expression of vascular endothelial growth factor. J Biol Chem 271: 736–741.
Dirix, L. Y., Vermeulen, P. B., Pawinski, A., Prove, A., Benoy, I., de Pooter, C., Martin, M. & van Oosterom, A. T. (1997). Elevated levels of the angiogenic cytokines basic fibroblast growth factor and vascular endothelial growth factor in sera of cancer patients. Br J Cancer 76: 238–243.
de Jong, J. S., van Diest, P. J., van der Valk, P. & Baak, J. P. A. (1998). Expression of growth factors, growth inhibiting factors and their receptors in invasive breast cancer. I: An inventory in search of autocrine and paracrine loops. J Pathol 184: 44–52.
de Vries, C., Escobedo, J. A., Ueno, H., Houck, K., Ferrara, F. & Williams, L. T. (1992). The fms-like tyrosine kinase, a receptor for vascular endothelial growth factor. Science 255: 989–991.
Eppenberger, U., Kueng, W., Schlaeppi, J. M., Roesel, J. L., Benz, C., Mueller, H., Matter, A., Zuber, M., Luescher, K., Litschgi, M., Schmitt, M., Foekens, J. A. & Eppenberger-Castori, S. (1998). Markers of tumor angiogenesis and proteolysis independently define high- and low-risk subsets of node-negative breast cancer patients. J Clin Oncol 16: 3129–3136.
Ergun, S., Kilic, N., Fiedler, W. & Mukhopadhyay, A. K. (1997). Vascular endothelial growth factor and its receptors in normal human testicular tissue. Mol Cell Endocrinol 131: 9–20.
Ferrara, N. & Davis-Smyth, T. (1997). The biology of vascular endothelial growth factor. Endo Rev 18: 4–25.
Folkman, J. (1990). What is the evidence that tumors are angiogenesis dependent?. J Natl Cancer Inst 82: 4–6.
Fukamara, D., Xavier, R., Sugiura, T., Chen, Y., Park, E-C, Lu, N., Selig, M., Nielsen, G., Taksir, T., Jain, R. K. & Seed, B. (1998). Tumor induction of VEGF promoter activity in stromal cells. Cell 94: 715–725.
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.
Green, A. R., Green, V. L., White, M. C. & Speirs, V. (1997). Expression of cytokine messenger RNA in normal and neoplastic human breast tissue: identification of interleukin-8 as a potential regulatory factor in breast tumours. Int J Cancer 72: 937–941.
Green, V. L., Atkin, S. L., Speirs, V., Jeffreys, R. V., Landolt, A. M., Mathew, B., Hipkin, L. & White, M. C. (1996). Cytokine expression in human anterior pituitary adenomas. Clin Endo 45: 179–185.
Hewett, P. W. & Murray, J. C. (1996). Coexpression of flt-1, flt-4 and KDR in freshly isolated and cultured human endothelial cells. Biochem Biophys Res Commun 221: 697–702.
Hlatky, L., Tsionou, C., Hahnfeldt, P. & Coleman, C. N. (1994). Mammary fibroblasts may influence breast tumour angiogenesis via hypoxia-induced vascular endothelial growth factor up-regulation and protein expression. Cancer Res 54: 6083–6086.
Jakeman, L. B., Winer, J., Bennett, G. L., Altar, C. A. & Ferrara, N. (1992). Binding sites for vascular endothelial growth factor are localised on endothelial cells in adult rat tissues. J Clin Invest 89: 244–253.
Kendall, R. L., Wang, G. & Thomas, K. A. (1996). Identification of a soluble form of the vascular endothelial growth factor receptor flt-1 and its heterodimerisation with KDR. Biochem Biophys Res Commun 226: 234–328.
Klagsbrun, M. & Soker, S. (1993). VEGF/VPS: the angiogenesis factor found?. Curr Biol 3: 699–702.
Koch, A. E., Polverini, P. J., Kunkel, S. L., Harlow, L. A., DiPietro, L. A., Elner, V. M., Elner, S. G. & Strieter, R. M. (1992). Interleukin-8 as a macrophage-derived mediator of angiogenesis. Science 258: 1798–1801.
Linderholm, B., Tavelin, B., Grankvist, K. & Henriksson, R. (1998). Vascular endothelial growth factor is of high prognostic value in node-negative breast carcinoma. J Clin Oncol 16: 3121–3128.
Lu, Q. & Brodie, A. (1996). Stimulation of the growth of MCF-7 and MDA-MB-468 breast cancer cells by vascular endothelial growth factor. Proc Am Assoc Cancer Res 37: 1499
Scott, P. A. E., Smith, K., Poulsom, R., De Benedetti, A., Bicknell, R. & Harris, A. L. (1998). Differential expression of vascular endothelial growth factor mRNA vs. protein isoform expression in human breast cancer and relationship to elF-4E. Br J Cancer 77: 2120–2128.
Seetharam, L., Gotoh, N., Maru, Y., Neufeld, G., Yamaguchi, S. & Shibuya, M. A. (1995). A unique signal transduction pathway for the flt-1 tyrosine kinase receptor, a receptor for vascular endothelial growth factor. Oncogene 10: 135–147.
Speirs, V., Green, A. R. & White, M. C. (1996a). Collagenase III: a superior enzyme for complete disaggregation and improved viability of normal and malignant human breast tissue. In Vitro Cell Dev Biol 32: 72–74.
Speirs, V., Green, A. R. & White, M. C. (1996b). A comparative study of cytokine gene transcripts in normal and malignant breast tissue and primary cell cultures derived from the same tissue samples. Int J Cancer 66: 551–556.
Speirs, V., Green, A. R., Walton, D. S., Kerin, M. J., Carleton, P. J., Fox, J. N., Desai, S. B. & Atkin, S. L. (1998). Short-term primary culture of epithelial cells derived from breast tumours. Br J Cancer 78: 1412–1429.
Terman, B. I., Dougher-Vermazen, M., Carrion, M. E., Dimitrov, D., Armellino, D. C., Gospodarowicz, D. & Bohlen, P. (1992). Identification of the KDR tyrosine kinase as a receptor for vascular endothelial growth factor. Biochem Biophys Res Commun 187: 1579–1586.
Tischer, E., Mitchell, R., Hartman, T., Silva, M., Gospodarowicz, D., Fides, J. C. & Abraham, J. A. (1991). The human gene for vascular endothelial growth factor. Multiple protein forms are encoded through alternative exon splicing. J Biol Chem 266: 11947–11954.
Toi, M., Kashitani, L. & Tominaga, T. (1993). Tumour angiogenesis is an independent prognostic indicator in primary breast carcinoma. Int J Cancer 55: 371–374.
Toi, M., Kondo, S., Suzuki, H., Yamamoto, Y., Inada, K., Imazawa, T., Taniguchi, T. & Tominaga, T. (1996). Quantitative analysis of vascular endothelial growth factor in primary breast cancer. Cancer 77: 1101–1106.
Waltenberger, J., Claesson-Welch, L., Siegbahn, A., Shibuya, M. & Heldin, C-H (1994). Different signal transduction properties of KDR and Flt-1, two receptors for vascular endothelial growth factor. J Biol Chem 269: 26988–26955.
Yoshiji, H., Gomez, D. E., Shibuya, M. & Thorgeirsson, U. P. (1996). Expression of vascular endothelial growth factor, its receptor, and other angiogenic factors in human breast cancer. Cancer Res 56: 2013–2016.
Weidner, N., Semple, J. P., Welch, W. R. & Folkman, J. (1991). Tumour angiogenesis and metastasis: correlation in invasive breast cancer. N Engl J Med 324: 1–8.
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
Speirs, V., Atkin, S. Production of VEGF and expression of the VEGF receptors Flt-1 and KDR in primary cultures of epithelial and stromal cells derived from breast tumours. Br J Cancer 80, 898–903 (1999). https://doi.org/10.1038/sj.bjc.6690438
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bjc.6690438
Keywords
This article is cited by
-
Effects of bisphenol A and 17β-estradiol on vascular endothelial growth factor A and its receptor expression in the non-cancer and cancer ovarian cell lines
Cell Biology and Toxicology (2015)
-
Exploiting the tumor microenvironment in the development of targeted cancer gene therapy
Cancer Gene Therapy (2009)
-
Expression of VEGF and VEGFR2 in tumors during neoadjuvant therapy of patients with breast cancer
Bulletin of Experimental Biology and Medicine (2008)
-
Phosphorylated KDR expression in endometrial cancer cells relates to HIF1α/VEGF pathway and unfavourable prognosis
Modern Pathology (2006)
-
Activation of mitogenic pathways and sensitization to estrogen-induced apoptosis: two independent characteristics of tamoxifen-resistant breast cancer cells?
Breast Cancer Research and Treatment (2006)