Specific expression of matrix metalloproteinases 1, 3, 9 and 13 associated with invasiveness of breast cancer cells in vitro.
Several matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) were studied in highly invasive (MDA-MB-231) and slightly invasive (MCF-7, T47D, BT-20) breast cancer cell lines. Investigations were carried out at the protein level and/or at the mRNA level, either in cells cultured as monolayers on plastic, or in cells seeded on a thin layer of Matrigel basement membrane matrix. Analysis of MMP expression by RT-PCR showed expression of MMP-1. MMP-3, and MMP-13 in highly invasive MDA-MB-231 cells, but not in slightly invasive cell lines. The extracellular secretion of MMP-1 and MMP-3 by MDA-MB 231 cells could be also shown by ELISA. TIMP-1 and TIMP-2 mRNAs were found in all cell lines, however, the extracellular secretion of both TIMPs was much higher in MDA-MB-231 cells than in the other cell lines. When the cells were cultured on Matrigel matrix, MMP-9 expression was induced in MDA-MB-231 cells only, as assessed by RT-PCR and zymography experiments. The invasive potential of MDA-MB-231 cells evaluated in vitro through Matrigel was significantly inhibited by the MMP inhibitor BB-2516, by 25% and 50% at the concentrations of 2 x 10(-6) M and 10(-5) M, respectively. In conclusion, our data show that highly invasive MDA-MB-231 cells but not slightly invasive T47D, MCF-7 and BT-20 cells express MMP-1, MMP-3, MMP-9 and MMP-13. MMP-9 which is specifically up-regulated by cell contact to Matrigel, may play a key role in the invasiveness of MDA-MB-231 cells through basement membranes.
Laboratoire de Biochimie, H pital Claude Huriez, Lille, France.
M
M
Balduyck
F
F
Zerimech
V
V
Gouyer
R
R
Lemaire
B
B
Hemon
G
G
Grard
C
C
Thiebaut
V
V
Lemaire
E
E
Dacquembronne
T
T
Duhem
A
A
Lebrun
M J
MJ
Dejonghe
G
G
Huet
D001483:Base Sequence; D001485:Basement Membrane / Q000201:enzymology; D001943:Breast Neoplasms / Q000201:enzymology* / Q000473:pathology; D003094:Collagen; D017931:DNA Primers; D004338:Drug Combinations; D004797:Enzyme-Linked Immunosorbent Assay; D006877:Hydroxamic Acids / Q000494:pharmacology; D007797:Laminin; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D011509:Proteoglycans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019714:Tissue Inhibitor of Metalloproteinases / Q000378:metabolism; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D004338:Drug Combinations; D006877:Hydroxamic Acids; D007797:Laminin; D061965:Matrix Metalloproteinase Inhibitors; D011509:Proteoglycans; D019714:Tissue Inhibitor of Metalloproteinases; C059485:matrigel; D003094:Collagen; C100342:marimastat; D020782:Matrix Metalloproteinases