Abstract
The expression of smooth muscle (SM) and non-muscle (NM) myosin heavy chain (MyHC) isoforms has been studied in fibroblastic cells of different fibrotic lesions (hypertrophic scars, Dupuytren's nodules and stromal reaction to mammary carcinoma) and in epithelial cells of non-neoplastic and neoplastic mammary glands, using anti-myosin antibodies in immunofluorescence and Western blotting. Two antibodies were specific for SM-MyHC isoforms (SM1 and SM2) and three antibodies were directed against different sequences of NM-MyHC isoforms. Myofibroblasts containing SM-MyHC were present in a variable number of cases of the different lesions: 1 of 11 hypertrophic scars, 3 of 9 Dupuytren' s nodules and 20 of 25 breast cancers. The distribution of NM-MyHC sequences recognized by our antibodies was heterogeneous in fibroblasts from normal dermis and mammary stroma, but became homogeneous in myofibroblasts from all the pathological conditions examined. Moreover, the expression of these MyHC sequences differed in normal mammary epithelium when compared with invasive carcinoma. These results show that cellular modulation from fibroblast to myofibroblast may be accompanied by the appearance of SM-MyHC and is characterized by a uniform expression of MyHC of NM type, and that tumour progression in mammary epithelial cells may be paralleled by the disappearance of a specific NM-MyHC sequence. This suggests that MyHC modulation participates in the process of fibrosis as well as in the process of malignant epithelial transformation.
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Babij P, Zhao J, White S, Woodcock-Mitchell J, Mitchell J, Absher M, Baldor L, Periasamy M, Low RB (1993) Smooth muscle myosin regulation by serum and cell density in cultured rat lung connective tissue cells. Am J Physiol 265:L17-L32
Benzonana G, Skalli O, Gabbiani G (1988) Correlation between the distribution of smooth muscle or non muscle myosins and α-smooth muscle actin in normal and pathological soft tissues. Cell Motil Cytoskeleton 11:260–274
Borrione AC, Zanellato AMC, Scannapieco G, Pauletto P, Sartore S (1989) Myosin heavy-chain isoforms in adult and developing rabbit vascular smooth muscle. Eur J Biochem 183:413–417
Borrione AC, Zanellato AMC, Giuriato L, Scannapieco G, Pauletto P, Sartore S (1990) Non muscle and smooth muscle myosin isoforms in bovine endothelial cells. Exp Cell Res 190:1–10
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Buoro S, Ferrarese P, Chiavegato A, Roelofs M, Scatena M, Pauletto P, Passerini-Glazel G, Pagano F, Sartore S (1993) Myofibroblast-derived smooth muscle cells during remodelling of rabbit urinary bladder wall induced by partial outflow obstruction. Lab Invest 69:589–602
Chiavegato A, Scatena M, Roelofs M, Ferrarese P, Pauletto P, Passerini-Glazel G, Pagano F, Sartore S (1993) Cytoskeletal and cytocontractile protein composition of smooth muscle cells in developing and obstructed rabbit bladder. Exp Cell Res 207:310–320
Clark RAF (1993) Regulation of fibroplasia in cutaneous wound repair. Am J Med Sci 306:42–48
Dalla Libera L, Sartore S (1981) Immunological and biochemical evidence for atrial-like isomyosin in thyrotoxic rabbit ventricle. Biochem Biophys Acta 669:84–92
Darby I, Skalli O, Gabbiani G (1990) α-Smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Invest 63:21–29
Desmoulière A, Rubbia-Brandt L, Grau G, Gabbiani G (1992) Heparin induces α-smooth muscle actin expression in cultured fibroblasts and in granulation tissue myofibroblasts. Lab Invest 67:716–726
Desmoulière A, Geinoz A, Gabbiani F, Gabbiani G (1993) Transforming growth factor-β1 induces α-smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent and growing cultured fibroblasts. J Cell Biol 122:103–111
Eddinger TJ, Murphy RA (1991) Developmental changes in actin and myosin heavy chain isoform expression in smooth muscle. Arch Biochem Biophys 284:232–237
Eddy RJ, Petro AJ, Tomasek JJ (1988) Evidence for the non-muscle nature of the “myofibroblast” of granulation tissue and hypertrophic scar. An immunofluorescence study. Am J Pathol 130:252–260
Ehrlich HP, Desmoulière A, Diegelmann RF, Cohen IK, Compton CC, Garner WL, Kapanci Y, Gabbiani G (1994) Morphological and immunochemical differences between keloid and hypertrophic scar. Am J Pathol 145:105–113
Fabra A, Nakajima M, Bucana CD, Fidler IJ (1992) Modulation of the invasive phenotype of human colon carcinoma cells by organ specific fibroblasts of nude mice. Differentiation 52:101–110
Frid MG, Printseva OY, Chiavegato A, Faggin E, Scatena M, Koteliansky VE, Pauletto P, Glukhova MA, Sartore S (1993) Myosin heavy-chain isoform composition and distribution in developing and adult human aortic smooth muscle. J Vasc Res 30:279–292
Giuriato L, Borrione AC, Zanellato AMC, Tonello M, Scatena M, Scannapieco G, Pauletto P, Sartore S (1991) Aortic intimal thickening and myosin isoform expression in hyperthyroid rabbits. Arterioscler Thromb 11:1376–1389
Giuriato L, Scatena M, Chiavegato A, Tonello M, Scannapieco G, Pauletto P, Sartore S (1992) Non-muscle myosin isoforms and cell heterogeneity in developing rabbit vascular smooth muscle. J Cell Sci 101:233–246
Grinnel F (1994) Fibroblasts, myofibroblasts, and wound contraction. J Cell Biol 124:401–404
Kawamoto S, Adelstein RS (1987) Characterization of myosin heavy chains in cultured aorta smooth muscle cells. A comparative study. J Biol Chem 262:7282–7288
Kelly CA, Takahashi M, Yu JH, Adelstein RS (1993) An insert of seven aminoacids confers functional differences between smooth muscle myosins from the intestine and vasculature. J Biol Chem 268:12|848–12|854
Kimura A, Nakashima S, Uda T, Ikeda H, Yasuda S, Tsuji T, Matsumura S (1993) Heavy-chain isoforms of non-muscle myosin in human tissues. Eur J Biochem 213:5–10
Komuro T (1990) Re-evaluation of fibroblasts and fibroblast-like cells. Anat Embryol 182:103–112
Kuro-o M, Nagai R, Tsuchimochi H, Katoh H, Yazaki Y, Ohkubo A, Takaku F (1989) Developmentally regulated expression of vascular smooth muscle myosin heavy chain isoforms. J Biol Chem 264:18|272–18|275
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lazard D, Sastre X, Frid MG, Glukhova MA, Thiery JP, Koteliansky VE (1993) Expression of smooth muscle-specific proteins in myoepithelium and stromal myofibroblasts of normal and malignant human breast tissue. Proc Natl Acad Sci USA 90: 999–1003
Lenette DA (1978) An improved mounting medium for immunofluorescence microscopy. Am J Clin Pathol 69:647–648
Liotta LA, Steeg PS, Stetler-Stevenson WG (1991) Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell 64:327–336
Murakami N, Elzinga M (1992) Immunohistochemical studies on the distribution of cellular myosin II isoforms in brain and aorta. Cell Motil Cytoskeleton 22:281–295
Osborn M, Weber K (1982) Intermediate filaments: cell-type-specific markers in differentiation and pathology. Cell 31:303–306
Pittet B, Rubbia-Brandt L, Desmoulière A, Sappino AP, Roggero P, Guerret S, Grimaud JA, Lacher R, Montandon D, Gabbiani G (1994) Effect of γ-interferon on the clinical and biologic evolution of hypertrophic scars and Dupuytren's disease: an open pilot study. Plast Reconstr Surg 93:1224–1235
Pollard TD, Thomas SM, Niederman R (1974) Human platelet myosin. I. Purification by a rapid method applicable to other nonmuscle cells. Anal Biochem 60:258–266
Redowicz MJ, Sobieszek A, Strzelecka-Golaszewska H (1990) Conformational transitions within the head and at the head-rod junction in smooth muscle myosin studied with a limited proteolysis method. Eur J Biochem 192:601–608
Ronnov-Jessen L, Petersen OW (1993) Induction of α-smooth muscle actin by transforming growth factor-β1 in quiescent human breast gland fibroblasts. Lab Invest 68:696–707
Rubbia-Brandt L, Sappino AP, Gabbiani G (1991) Locally applied GM-CSF induces the accumulation of α-smooth muscle actin containing myofibroblasts. Virchows Arch [B] 60:73–82
Sappino AP, Schürch W, Gabbiani G (1990) Differentiation repertoire of fibroblastic cells: expression of cytoskeletal proteins as marker of phenotypic modulations. Lab Invest 63:144–161
Schmitt-Gräff A, Desmoulière A, Gabbiani G (1994) Heterogeneity of myofibroblast phenotypic features: an example of fibroblastic cell plasticity. Virchows Arch 425:3–24
Schürch W, Skalli O, Gabbiani G (1990) Cellular biology of Dupuytren's disease. In: McFerland RM, McGrouther DA, Flint MH (eds) Dupuytren's disease. Churchill Livingstone, London, pp 31–47
Schürch W, Seemayer TA, Gabbiani G (1992) Myofibroblast. In: Sternberg SS (ed) Histology of pathologists. Raven Press, New York, pp 109–144
Sellers JR, Harvey EV (1984) Localization of light-chain binding site on smooth muscle myosin revealed by light-chain overlay of sodium dodecyl sulphate-polyacrylamide electrophoretic gels. J Biol Chem 259:14|203–14|207
Skalli O, Gabbiani G (1990) The biology of the myofibroblast and its relation to the development of soft tissue and epithelial tumours. In:Fletcher CDM, McKee PH (eds) Pathobiology of soft tissue tumours. Churchill Livingstone, Edinburgh, pp 83–103
Skalli O, Ropraz P, Trzeciak A, Benzonana G, Gillessen D, Gabbiani G (1986) A monoclonal antibody against α-smooth muscle actin: a new probe for smooth muscle differentiation. J Cell Biol 103:2787–2796
Skalli O, Schürch W, Seemayer T, Lagacé R, Montandon D, Pittet B, Gabbiani G (1989) Myofibroblasts from diverse pathologic settings are heterogeneous in their content of actin isoforms and intermediate filament proteins. Lab Invest 60:275–285
Takahashi M, Kawamoto S, Adelstein RS (1992) Evidence for inserted sequences in the head region of nonmuscle myosin specific to the nervous system. Cloning of the cDNA encoding the myosin heavy chain-B isoform of vertebrate nonmuscle myosin. J Biol Chem 267:17|864–17|871
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Vandekerckhove J, Weber K (1978) At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal tryptic peptide. J Mol Biol 126:783–802
Van denHooff A (1988) Stromal involvement in malignant growth. Adv Cancer Res 50:159–196
Vracko R, Thorning D (1993) Myofibroblasts and smooth muscle cells in human myocardial scars: possible origins and inductive factors. Cardiovasc Pathol 2:207–213
Vyalov S, Desmoulière A, Gabbiani G (1993) GM-CSF-induced granulation tissue formation: relationships between macrophage and myofibroblast accumulation. Virchows Arch [B] 63:231–239
Whalen RG, Sell SM, Brutler-Browne GS, Schawartz K, Bouveret P, Pinset-Harstrom I (1981) Three myosin heavy chain isoenzymes appear sequentially in rat muscle development. Nature 292:805–809
Zanellato AMC, Borrione AC, Giuriato L, Tonello M, Scannapieco G, Pauletto P, Sartore S (1990) Myosin isoforms and cell heterogeneity in vascular smooth muscle. I. Developing and adult bovine aorta. Dev Biol 141:431–446
Zanellato AMC, Borrione AM, Tonello M, Scannapieco G, Pauletto P, Sartore S (1990) Myosin isoform expression and smooth muscle cell heterogeneity in normal and atheriosclerotic rabbit aorta. Arteriosclerosis 10:996–1009
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Chiavegato, A., Sartore, S., D'Amore, E. et al. Expression of myosin heavy chain isoforms in mammary epithelial cells and in myofibroblasts from different fibrotic settings during neoplasia. Vichows Archiv A Pathol Anat 426, 77–86 (1995). https://doi.org/10.1007/BF00194701
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DOI: https://doi.org/10.1007/BF00194701