Abstract
Melanoma invasion is a complex multi stage process involving changes to the cell/extracellular matrix (ECM) and cell/cell interactions. We have previously shown using an in vitro model of reconstructed human skin (consisting of human dermis with a basement membrane [BM] and populated with human skin cells) that some melanoma cells (HBL cell line) invade more actively in the presence of adjacent normal skin cells. The aim of the present study was to further investigate the relationship between melanoma cells, skin cells and ECM proteins during melanoma cell invasion through reconstructed skin, extending this to a study of three melanoma cell lines. We also examined whether such cell/cell induced invasion is due to increased expression and activation of matrix-metalloproteinase-2 (MMP-2) and MMP-9, or due to increases in general protease activity for keratinocytes, fibroblasts or melanoma lines. Addition of skin cells dramatically altered the invasive behaviour of the three metastatic melanoma cell lines (HBL, C8161 and A375SM) used; they increased the invasive ability of HBLs which were unable to invade on their own; they potentiated the invasion of C8161 cells which were invasive in their own right, but reduced the invasion of A375-SM cells which were aggressive invaders in the absence of skin cells. Latent forms of MMP-2, and MMP-9, were clearly expressed by the normal skin cells whereas all three melanoma lines weakly expressed these proteases. Fibroblast and keratinocyte MMPs were activated specifically by culture on type I collagen and on dermis which retained an intact basement membrane. These findings demonstrate that while there is an active communication between melanoma cells and adjacent skin cells, the invasive process is dictated by the melanoma cells and not the skin cells. However, activation of skin cell derived MMPs may play an important role in facilitating invasion by particular melanoma phenotypes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Blackwood MA, Holmes R, Synnestvedt M et al. Multiple primary melanoma revisited. Cancer 2002; 94 (8): 2248–55.
Li G, Herlyn M. Dynamics of intercellular communication during melanoma development. Mol Med Today 2000; 6: 163–9.
Halaban R, Langdon R, Birchall N et al. Basic fibroblast growth factor from human keratinocytes is a natural mitogen for melanocytes. J Cell Biol 1988; 107: 1611–9.
Yaar M, Grossman K, Eller Met al. Evidence for nerve growth factormediated paracrine effects in human epidermis. J Cell Biol 1991; 115 (3): 821–8.
Imokawa G, Yada Y, Miyagishi M. Endothelins secreted from human keratinocytes are intrinsic mitogens for human melanocytes. J Biol Chem 1992; 267 (34): 24675–80.
Rubin JS, Bottaro DP, Chedid M et al. Keratinocyte growth factor. Cell Biol Int 1995; 19 (5): 399–411.
Gilchrest BA, Albert LS, Karassik RL et al. Substrate influences human epidermal melanocyte attachment and spreading in vitro. In Vitro Cell Dev Biol 1985; 21 (2): 114–20.
Morelli JG, Yohn JJ, Zekman T et al. Melanocyte movement in vitro: role of matrix proteins and integrin receptors. J Invest Dermatol 1993; 101: 605–8.
Hara M, Yaar M, Tang A et al. Role of integrins in melanocyte attachment and dendricity. J Cell Sci 1994; 107: 2739–48.
Hedley S, Gawkrodger DJ, Weetman AP et al. Investigation of the influence of extracellular matrix proteins on normal human melanocyte morphology and melanogenic activity. Br J Dermatol 1996; 135: 888–97.
Hedley SJ, Layton C, Heaton M et al. Fibroblasts play a regulatory role in the control of pigmentation in reconstructed human skin from skin types I and II. Pigment Cell Res 2002; 15 (1): 49–56.
Guo M, Toda K, Grinnell F. Activation of human keratinocyte migration on type I collagen and fibronectin. J Cell Sci 1990; 96 (2): 197–205.
Edward M, MacKie RM. Cell-cell and cell-extracellular matrix interactions during melanoma cell invasion and metastasis. Melanoma Res 1993; 3: 227–34.
Hofmann UB, Westphal JR, van Muijen GNP et al. Matrix Metalloproteinases in human melanoma. J Invest Dermatol 2000; 115: 337–44.
Bodey B, Bodey B Jr, Siegel SE et al. Matrix Metalloproteinase expression in malignant melanomas: Tumor-extracellular matrix interactions in invasion and metastasis. In Vivo 2001; 15: 57–64.
Väisänen A, Tuominen H, Kallioinen M et al. Matrix metalloproteinase-2 (72 kD type IV collagenase) expression occurs in the early stage of human melanocytic tumour progression and may have prognostic value. J Pathol 1996; 180: 283–9.
Cornil I, Theodorescu D, Man S et al. Fibroblast cell interactions with human melanoma cells affect tumour cell growth as a function of tumour progression. Proc Natl Acad Sci USA 1991; 88: 6028–32.
Benbow U, Schoenermark MP, Mitchell TL et al. A novel host/tumor cell interaction activates matrix metalloproteinase-1 and mediates invasion through type I collagen. J Biol Chem 1999; 274 (36): 25371–8.
Edward M. Melanoma cell-derived factors stimulate glycosaminoglycan synthesis by fibroblasts cultured as monolayers and within contracted collagen lattices. Br J Dermatol 2001; 144: 465–70.
Bessou-Touya S, Morichon F, Surleve-Bazeille J-E et al. An ex vivo study of congenital pigmented nevi in epidermal reconstructs. Pigment Cell Res 1999; 12: 164–74.
Meier F, Nesbit M, Hsu M-Y et al. Human melanoma progression in skin reconstructs: biological significance of bFGF. Am J Pathol 2000; 156: 193–200.
Dekker SK, Van Doorn R, Kempenaar J et al. Skin equivalent: An attractive model to evaluate early melanoma metastasis. Melanoma Res 2000; 10: 127–40.
Bechetoille N, Haftek M, Staquet M-J et al. Penetration of human metastatic melanoma cells through an authentic dermal-epidermal junction is associated with dissolution of native collagen types IV and VII. Melanoma Res 2000; 10: 427–34.
Eves P, Layton C, Hedley S et al. Characterisation of an in vitro model of human melanoma invasion based on reconstructed human skin. Br J Dermatol 2000; 142: 210–22.
Chakrabarty KH, Dawson RA, Harris P et al. Development of autologous human dermal-epidermal composites based on sterilised human allo-dermis for clinical use. Br J Dermatol 1999; 141: 811–23.
Ghanem G, Comunale G, Libert A et al. Evidence for alphamelanocyte stimulating hormone (?-MSH) receptors on human malignant melanoma cells. Int J Cancer 1988; 41: 248–55.
Giard DJ, Aaronson SA, Todaro GJ et al. Iatn vitro cultivation of human tumours: establishment of cell lines derived from a series of solid tumours. J Natl Cancer Inst 1973; 51: 1417–23.
Bregman MD, Meyskens FL Jr. In vitro modulation of human and murine melanoma growth by prostanoid analogues. Prostaglandins 1983; 26: 449–56.
Hendrix MJ, Seftor EA, Chu YW et al. Coexpression of vimentin and keratins by human melanoma tumour cells: correlation with invasive and metastatic potential. J Natl Cancer Inst 1992; 84 (3): 165–74.
Ralston DR, Layton C, Dalley AJ et al. The requirement for basement membrane antigens in the production of human epidermal/dermal composites in vitro. Br J Dermatol 1999; 140: 605–15.
Welch DR, Bisi JE, Miller BE et al. Characterisation of a highly invasive and spontaneously metastatic human malignant melanoma cell line. Int J Cancer 1991; 47: 227–37.
Kähäri V-M, Saarialho-Kere U. Matrix Metallproteinases in skin. Exp Dermatol 1997; 6: 199–213.
Laplante AF, Germain L, Auger FA et al. Mechanisms of wound reepithelialisation: Hints from a tissue-engineered reconstructed skin to long-standing questions. FASEB J 2001; 15 (13): 2377–89.
Grayson R, Douglas CWI, Heath J et al. Activation of human matrixmetalloproteinase-2 by gingival crevicular fluid and porphyromonas gingivalis: J Clin Periodontol2003; 30(6): 542–50.
Banerjee SS, Harris M. Morphological and immunophenotypic variations in malignant melanoma. Histopathology 2000; 36: 387–402.
Cooke ME, Sakai T, Mosher DF. Contraction of collagen matrices mediated by alpha-2 beta 1A and alpha (v) beta3 integrins. J Cell Sci 2000: 113: 2375–83.
Chakrabarty KH, Heaton M, Dalley AJ et al. Keratinocyte-driven contraction of reconstructed human skin. Wound Rep Regen 2001;9 (2): 95–106.
Bullen EC, Longaker MT, Updike DL et al. Tissue inhibitor of metalloproteinase-1 is decreased and activated gelatinases are increased in chronic wounds. J Invest Dermatol 1995; 104: 236–40.
Zhu N, Haycock JW, Eves PC et al. ?-MSH attenuates TNF-? stimulated integrin expression and cell adhesion in human melanoma cells. J Invest Dermatol 2002; 119(5): 1165–71.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Eves, P., Katerinaki, E., Simpson, C. et al. Melanoma invasion in reconstructed human skin is influenced by skin cells – investigation of the role of proteolytic enzymes. Clin Exp Metastasis 20, 685–700 (2003). https://doi.org/10.1023/B:CLIN.0000006824.41376.b0
Issue Date:
DOI: https://doi.org/10.1023/B:CLIN.0000006824.41376.b0