Summary
Myofibroblasts from human breast carcinomas were identified and experimentally generated in culture, and a possible function was examined. The frequency ofα-smooth muscle actin immunoreactive cells was evaluated as a measure of myofibroblast differentiation in primary culture. Few or noα-smooth muscle actin-positive stromal cells (6.1 ± 8.4%) were identified in primary cultures from normal breast tissue (n=9). In contrast, high frequencies (68.8 ± 15.1%) were observed in primary cultures from carcinomas (n=19). The frequencies of myofibroblasts in primary cultures were almost identical to those obtained in the corresponding cryostat sections (69.1 vs. 68.8%). A possible precursor cell to the myofibroblast was looked for among typical fibroblasts and vascular smooth muscle cells. Purified blood vessels containing both fibroblasts and vascular smooth muscle cells were embedded in collagen gel and incubated with medium conditioned by breast epithelial cells. Fibroblasts rather than smooth muscle cells were recruited from the blood vessels. In medium conditioned by carcinoma cell lines or in co-cultures of carcinoma cell lines and purified fibroblasts,α-smooth muscle actin and the typical myofibroblast phenotype were induced in otherwiseα-smooth muscle actin-negative fibroblasts. The effect of myofibroblasts on cellular movement—essential to neoplastic cells—was analyzed. Spontaneous motility of tumor cells (MCF-7) was entirely suppressed in a collagen gel assay. Under these conditions tumor cell motility was selectively mediated by direct cell-to-cell interaction between tumor cells and myofibroblasts. Under chemically defined conditions, interaction was dependent on the presence of plasminogen. Anti-plasminogen, soybean trypsin inhibitor, and anti-fibronectin partly neutralized the effect of plasminogen. It is concluded that elements of myofibroblast differentiation and function may be studied in culture.
Similar content being viewed by others
References
Adams, E. F.; Newton, C. J.; Braunsberg, H., et al. Effects of human breast fibroblasts on growth and 17β-estradiol dehydrogenase activity of MCF-7 cells in culture. Breast Cancer Res. Treat. 11:165–172; 1988.
Barsky, S. H.; Roa, C. N.; Grotendorst, G. R., et al. Increased content of type V collagen in desmoplasia of human breast carcinoma. Am. J. Pathol. 108:276–283; 1982.
Basset, P.; Bellocq, J. P.; Wolf, C., et al. A novel metalloproteinase gene specifically expressed in stromal cells of breast carcinomas. Nature 348:699–704; 1990.
Briand, P.; Lykkesfeldt, A. Long-term cultivation of a human breast cancer cell line, MCF-7, in a chemically defined medium. Effect of estradiol. Anticancer Res. 6:85–90; 1986.
Briand, P.; Petersen, O. W.; van Deurs, B. A new diploid nontumorigenic human breast epithelial cell line isolated and propagated in chemically defined medium. In Vitro Cell. Dev. Biol. 23:181–188; 1987.
Bronzert, D. A.; Pantazis, P.; Antoniades, H. N., et al. Synthesis and secretion of platelet-derived growth factor by human breast cancer cell lines. Proc. Natl. Acad. Sci. USA 84:5763–5767; 1987.
Camps, J. L.; Chang, S.-M.; Hsu, T. C., et al. Fibroblast-mediated acceleration of human epithelial tumor growth in vivo. Proc. Natl. Acad. Sci. USA 87:75–79; 1990.
Chamley, J. H.; Gröschel-Steward, U.; Campbell, G. R., et al. Distinction between smooth muscle, fibroblasts and endothelial cells in culture by use of fluoresceinated antibodies against smooth muscle actin. Cell Tissue Res. 177:445–457; 1977.
Chamley-Campbell, J. H.; Campbell, G. What controls smooth muscle phenotype? Atherosclerosis 40:347–357; 1981.
Chiquet-Ehrismann, R.; Kalla, P.; Pearson, C. A. Participation of tenascin and transforming growth factor-β in reciprocal epithelial-mesenchymal interactions of MCF7 cells and fibroblasts. Cancer Res. 49:4322–4325; 1989.
Chiquet-Ehrismann, R.; Kalla, P.; Pearson, C. A., et al. Tenascin interferes with fibronectin action. Cell 53:383–390; 1988.
Chiquet-Ehrismann, R.; Mackie, E. J.; Pearson, C. A., et al. Tenascin: an extracellular matrix protein involved in tissue interactions during fetal development and oncogenesis. Cell 47:131–139; 1986.
Darby, I.; Skalli, O.; Gabbiani, G.α-Smooth muscle muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab. Invest. 63:21; 1990.
Gleiber, W. E.; Schiffmann, E. Identification of a chemoattractant for fibroblasts produced by human breast carcinoma cell lines. Cancer Res. 44:3398–3402; 1984.
Grøndahl-Hensen, J.; Ralfkiær, E.; Kirkeby, L. T., et al. Localization of urokinase-type plasminogen activator in stromal cells in adenocarcinomas of the colon in humans. Am. J. Pathol. 138:111–117; 1991.
Guidry, C.; Hohn, S.; Hook, M. Endothelial cells secrete a factor that promotes fibroblast contraction of hydrated collagen gels. J. Cell Biol. 110:519–528; 1990.
Haslam, S. Z. Mammary fibroblast influence on normal mouse mammary epithelial cell responses to estrogen in vitro. Cancer Res. 46:310–316; 1986.
Klagsburn, M. Biosynthesis and storage of basic fibroblast growth factor (bFGF) by endothelial cells: implication for the mechanism of action of angiogenesis. In: Back, N.; Brewer, G. J.; Eijsvoogel, V. P., et al. eds. Growth factors and other aspects of wound healing: biological and clinical implications. New York: Arthur R. Liss; 1988:55–61.
Lippman, M. E.; Dickson, R. B.; Bates, S., et al. Autocrine and paracrine growth regulation of human breast cancer. Breast Cancer Res. Treat. 7:59–70; 1986.
Lippman, M. E.; Dickson, R. B.; Gelmann, E. P., et al. Growth regulatory peptide production by human breast carcinoma cells. J. Steroid Biochem. 30:53–61; 1988.
Mackie, E. J.; Halfter, W.; Liverani, D. Induction of tenascin in healing wounds. J. Cell Biol. 107:2757–2767; 1988.
McGrath, C. M. Augmentation of the response of normal mammary epithelial cells to estradiol by mammary stroma. Cancer Res. 43:1355–1360; 1983.
Mignatti, P.; Robbins, E.; Rifkin, D. B. Tumor invasion through the human amniotic membrane: requirement for a proteinase cascade. Cell 47:487–498; 1986.
Oda, D.; Gown, A. M.; Vande Berg, J. S., et al. The fibroblast-like nature of myofibroblasts. Exp. Mol. Pathol. 49:316–329; 1988.
Ohtani, H.; Sasano, N. Myofibroblasts and myoepithelial cells in human breast carcinoma. Virchow. Arch. A Path. Anat. Histol. 385:247–261; 1980.
Owens, g. K.; Loeb, A.; Gordon, D., et al. Expression of smooth muscle-specificα-isoaction in cultured vascular smooth muscle cells: relationship between growth and cytodifferentiation. J. Cell Biol. 102:343–352; 1986.
Pallesen, G.; Nielsen, S.; Celis, J. E. Characterization of a monoclonal antibody (BG3C8) that reacts with basal cells of stratified epithelia. Histopathology 11:591–601; 1987.
Petersen, O. W.; Hansen, S. H.; Laursen, I., et al. Effect of insulin on growth and expression of smooth muscle isoactin in human breast gland myoepithelial cells in a chemically defined culture system. Eur. J. Cell Biol. 50:500–509; 1989.
Petersen, O. W.; van Deurs, B. Preservation of defined phenotypic traits in short-time cultured human breast carcinoma derived epithelial cell. Cancer Res. 47:856–866; 1987.
Petersen, O. W.; van Deurs, B. Growth factor control of myoepithelialcell differentiation in cultures of human mammary gland. Differentiation 39:197–215; 1988.
Petersen, O. W.; van Deurs, B. Distinction between vascular smooth muscle cells and myoepithelial cells in primary monolayer cultures of human breast tissue. In Vitro Cell Dev. Biol. 25:259–266; 1989.
Petersen, O. W.; van Deurs, B.; Vang Nielsen, K., et al. Differential tumorigenecity of two autologous human breast carcinoma cell lines HMT-3909 S1 and HMT-3909 S8 established in serum-free medium. Cancer Res. 50:1–14; 1990.
Rosen, E. M.; Goldberg, I. D. Protein factors which regulate cell motility. In Vitro Cell. Dev. Biol. 25:1079–1087; 1989.
Rønnov-Jessen, L.; van Deurs, B.; Celis, J. E., et al. Smooth muscle differentiation in cultured human breast gland stromal cells. Lab. Invest. 63:532–543; 1990.
Sappino, A.-P.; Schürch, W.; Gabbiani, G. Biology of disease. Differentiation repertoire of fibroblastic cells: expression of cytoskeletal proteins as marker of phenotypic modulations. Lab. Invest. 63:144–161; 1990.
Sappino, A.-P.; Skalli, O.; Jackson, B., et al. Smooth-muscle differentiation in stromal cells of malignant and non-malignant breast tissues. Int. J. Cancer. 41:707–712; 1988.
Sato, Y.; Rifkin, D. B. Inhibition of endothelial cell movement by pericytes and smooth muscle cells: activation of a latent transforming growth factor-β1-like molecular by plasmin during co-culture. J. Cell Biol. 109:309–315; 1989.
Schürch, W.; Seemayer, T. A.; Lagacé, R. Stromal myofibroblasts in primary invasive and metastatic carcinomas. Virchows Arch. [A]. 391:125–139; 1981.
Seemayer, T. A.; Schürch, W.; Lagacé, R. Myofibroblasts in human pathology. Hum. Pathol. 12:491–492; 1981.
Singer, K. H.; Searce, R. M.; Tuck, D. T., et al. Removal of fibroblasts from human epithelial cell cultures with use of a complement fixing monoclonal antibody reactive with human fibroblasts and monocytes/macrophages. J. Invest. Dermatol. 92:166–170; 1989.
Skalli, O.; Gabbiani, G. The biology of the myofibroblast relationship to wound contraction and fibrocontractive diseases. In: Clark, R. A. F.; Henson, P. M., eds. The molecular and cellular biology of wound repair. New York: Plenum Publishing Corporation; 1988:373–402.
Skalli, O.; Pelte, M.-F.; Peclet, M.-C., et al.α-Smooth muscle actin, a differentiation marker of smooth muscle cells, is present in microfilamentous bundles of pericytes. J. Histochem. Cytochem. 37:315–321; 1989.
Skalli, O.; Ropraz, P.; Trzeciak, A., et al. A monoclonal antibody againstα-smooth muscle actin: a new probe for smooth muscle differentiation. J. Cell. Biol. 103:2787–2796; 1986.
Skalli, O.; Schürch, W.; Seemayer, T., et al. Myofibroblasts from diverse pathologic settings are heterogeneous in their content of actin isoforms and intermediate filament proteins. Lab. Invest. 60:275–285; 1989.
Tremblay, G. Stromal aspects of breast carcinoma. Exp. Mol. Pathol. 31:248–260; 1979.
Tsukada, T.; McNutt, M. A.; Ross, R., et al. HHF35, a muscle actinspecific monoclonal antibody II. Reactivity in normal, reactive and neoplastic human tissues. Am. J. Pathol. 127:389–402; 1987.
Tsukada, T.; Tippens, D.; Gordon, D., et al. HHF35, a muscle-actinspecific monoclonal antibody. Am. J. Pathol. 126:51–60; 1987.
Varani, J.; McKeever, P. E.; Fligiel, S. E. G., et al. Plasminogen activator production by human tumor cells: effect on tumor cell-extracellular matrix interactions. Int. J. Cancer. 40:772–777; 1987.
Welch, M. P.; Odland, G. F.; Clark, R. A. F. Temporal relationship of F-actin bundle formation, collagen and fibronectin matrix assembly, and fibronectin receptor expression to wound contraction. J. Cell Biol. 110:113–145; 1990.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rønnov-Jessen, L., van Deurs, B., Nielsen, M. et al. Identification, paracrine generation, and possible function of human breast carcinoma myofibroblasts in culture. In Vitro Cell Dev Biol - Animal 28, 273–283 (1992). https://doi.org/10.1007/BF02634244
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02634244