Inhibition of collagenolytic activity relates to quantitative reduction of invasionin vitro in a c-Ha-ras transfected glial cell line

Abstract

The function of proteases in brain tumor invasion is currently not well established. For tumors of epithelial and fibromatous origin collagenase production can enhance the invasive capacity of cells to penetrate basement membranes. We showed previously that a c-Ha-ras transformed glial cell line (CxT24neo3) invaded hamster brain tissue in vivo. These cells were also capable of invading reconstituted basement membrane and embryonic chick hearts in vitro. Since the histopathology of CxT24neo3 tumors mimics that of glioblastoma multiforme in humans, CxT24neo3 was used as the model in vitro for this type of brain tumor. Presently, we detected by zymogram analysis a gelatinase that was secreted by CxT24neo3 and that had an apparent molecular mass of 62 kD. To verify whether gelatinase affected invasion in vitro of these glial cells we determined the efficacy of a substrate specific collagenase inhibitor on invasion in vitro. GM6001 is a synthetic polypeptide that specifically occupies the substrate binding sites of metalloprotease. Since this drug did not show cytotoxicity, its specificity for metalloprotease is a valuable tool to evaluate the physiological function of these enzymes on invasion. We found that treatment of CxT24neo3 with GM6001 reduced the fraction of invading CxT24neo3 cells through reconstituted basement membrane. These data suggest that metalloproteases can stimulate brain tumor invasion.

Helicobacter pylori CagA induces a transition from polarized to invasive phenotypes in MDCK cells

“…In contrast, only 0.6% of control cells (n ϭ 1,890) penetrated the same matrix (t test P Ͻ 0.0001). We also investigated whether MMPs are involved in this invasive behavior by determining whether the number and size of CagA-induced invasive processes would respond to treatment with GM6001, a synthetic peptide inhibitor of MMPs (19). MDCK monolayers polarized on Matrigel-coated filters were pretreated with the MMP inhibitor before transfection.…”

Invasion by Matrix Metalloproteinase-Expressing Cells Is Important for Primitive Streak Formation in Early Chick Blastoderm

“…5 c, d, f). GM 6001 is a potent inhibitor of various MMPs in tumor invasion [Boghaert et al, 1994;Santibáñez et al, 2002;Yeh et al, 2006] and has been reported to inhibit fin regeneration in zebrafish [Bai et al, 2005]. The present study reveals that at least 3 kinds of MMPs continue to be localized in the area pellucida from the prestreak stage to the streak stage ( fig.…”

Echoes of a Distant Past: The cag Pathogenicity Island of Helicobacter pylori

“…When GM6001, a synthetic peptide inhibitor of MMPs (Boghaert et al 1994) is added to the matrix a four-times reduction was observed, suggesting that the process is MMPs dependent and supporting the notion that CagA induces invasive behavior in epithelial cells (Kitadai et al 2003;El-Etr et al 2004). Different domains of the CagA protein were expressed in polarized MDCK monolayers.…”