Modulation of invasive properties of murine squamous carcinoma cells by heterologous expression of cathepsin B and cystatin C.
Murine SCC-VII squamous carcinoma cells have the capacity to penetrate reconstituted basement membranes (Matrigel) in vitro. The invasion of Matrigel layers by SCC-VII cells was significantly reduced by E-64, a specific inhibitor of lysosomal cysteine proteinases. The cathepsin-B-selective E-64 derivative, CA-074, inhibited penetration of Matrigel by SCC-VII cells to the same extent, indicating a major role for this particular lysosomal enzyme in extracellular-matrix degradation during squamous-carcinoma-cell invasion. SCC-VII cells were stably transfected with a cDNA encoding human procathepsin B, in an attempt to modulate the invasive properties of the cell line. The transfected cells expressed the heterologous gene, secreted increased amounts of procathepsin B and displayed enhanced invasive potential. In vivo, the activity of cathepsin B is strictly regulated by endogenous inhibitors. SCC-VII cells were therefore also stably transfected with a cDNA encoding human cystatin C, the most potent cysteine-proteinase inhibitor in mammalian tissues. The expression of this transgene resulted in the production of active recombinant cystatin C and a pronounced reduction in Matrigel invasion. These studies demonstrate that the invasive properties of squamous-cell carcinomas can be changed by modulation of the balance between cathepsin B and its endogenous inhibitors, and provide further evidence for the involvement of this lysosomal cysteine proteinase in tumour invasion and metastasis.
Coulibaly
S
S
Zentrum für Angewandte Genetik, Universität für Bodenkultur, Vienna, Austria.
Schwihla
H
H
Abrahamson
M
M
Albini
A
A
Cerni
C
C
Clark
J L
JL
Ng
K M
KM
Katunuma
N
N
Schlappack
O
O
Glössl
J
J
Mach
L
L
D000818:Animals; D015152:Blotting, Northern; D002294:Carcinoma, Squamous Cell / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D002401:Cathepsin B / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D002633:Chemotaxis; D003094:Collagen / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000378:metabolism; D055316:Cystatin C; D015891:Cystatins / Q000096:biosynthesis* / Q000235:genetics; D015853:Cysteine Proteinase Inhibitors / Q000096:biosynthesis* / Q000235:genetics; D004338:Drug Combinations; D004792:Enzyme Precursors / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007797:Laminin / Q000378:metabolism; D051379:Mice; D009361:Neoplasm Invasiveness / Q000235:genetics; D011509:Proteoglycans / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
C525030:CST3 protein, human; C525031:Cst3 protein, mouse; D017077:Culture Media, Conditioned; D055316:Cystatin C; D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors; D004338:Drug Combinations; D004792:Enzyme Precursors; D007797:Laminin; D011509:Proteoglycans; D011994:Recombinant Proteins; C059485:matrigel; D003094:Collagen; C057832:procathepsin B; D002401:Cathepsin B