Fig. 8. GM6001 inhibits epimorphin-induced branching morphogenesis in a mammary
epithelial cell line. (A) Preclustered SCp2 cells were treated for 6 days in
collagen I gels with EGF, EGF+epimorphin (EPM) with or without GM6001. GM6001
inhibited branching induced by epimorphin. This inhibition was statistically
significant (***P
0.001). Other proteinase inhibitors
such as leupeptin, aprotinin, pepstatin and E64 did not show any inhibition of
branching (not shown). The mean±s.d. of three independent experiments
is shown. (B) Appearance of SCp2 cell clusters treated with EGF, EGF+EPM and
EGF+EPM+GM6001. (C) Epimorphin upregulates gelatinases A and B and Str1 in
SCp2 cells. SCp2 cells were treated for 48 hours with epimorphin or PBS as a
control. Conditioned medium was collected, concentrated and separated on
substrate gels. (D) Epimorphin regulates Str1 mRNA levels. RT-PCR analysis of
Str1 expression in SCp2 cells treated with PBS or EPM for 48 hours. Twenty,
23, 25, 30 and 35 cycles of amplification were performed for detection of Str1
and actin. While the level of actin was not different statistically in the PBS
and epimorphin-treated cultures (slightly higher in PBS in this experiment),
Str1 levels were clearly higher in epimorphin-treated cultures. Scale bars:
200 µm in A.