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Invasion of melanoma cells into dermal connective tissue in vitro : Evidence for an important role of cysteine proteases
Author(s) -
Dennhöfer Ralf,
Kurschat Peter,
Zigrino Paola,
Klose Anke,
Bosserhoff Anja,
van Muijen Goos,
Krieg Thomas,
Mauch Cornelia,
Hunzelmann Nicolas
Publication year - 2003
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.11255
Subject(s) - proteases , leupeptin , melanoma , connective tissue , extracellular matrix , matrix metalloproteinase , basement membrane , pathology , cysteine protease , cell culture , biology , chemistry , cancer research , microbiology and biotechnology , biochemistry , protease , enzyme , medicine , genetics
Invasion of melanoma cells into the dermal connective tissue is a major characteristic in the complex process of metastasis. Proteases play an important role in tumor cell invasion as these enzymes are able to degrade most components of the extracellular matrix (ECM), and thus enable cells to penetrate interstitial connective tissues and basement membranes. We developed an improved culture model that allows the detailed study of melanoma cell invasion in vitro . In this model, high (BLM) or low (530) invasive melanoma cells were seeded on the dermal side of dead deepidermized dermis (DDD) and cultured for 14 days at the air/liquid interface. The high invasive cells invaded the tissue, leading to dermal tumor formation, whereas the low invasive cells did not. Analysis of the enzymatic activity of gelatinases by in situ gelatin zymography at neutral pH revealed proteolysis only in those composites cultured with high invasive melanoma cells. Interestingly, in situ zymograms performed at more acidic conditions, favoring the activity of cysteine proteases, exhibited markedly enhanced and widespread gelatinolysis compared to neutral pH. Cysteine protease inhibitors (E‐64 and leupeptin) significantly reduced invasion of melanoma cells into these composites. These results indicate an important role of cysteine proteases for tumor invasion. © 2003 Wiley‐Liss, Inc.