Metalloproteinases are secreted by glioma cell line and induced VE-cadherin cleavage.

<p>(A) Conditioned media from Astrocytoma grade IV (LN229, LN) and Astrocytoma grade III (U87, U) cells lines were tested for protease activities using a zymography assay. Inhibition of protease activities by EDTA identified MMPs. (B) U87 (U) cell line media induced VE-cadherin cleavage from HUVECs (90 kDa fragment). Glioma cell line conditioned media was added to HUVEC confluent monolayer during two hours and HUVEC (H) conditioned media was analyzed for sVE content by western blot. The effect was impaired by broad spectrum MMPs inhibitor (GM6001, I). (C) Western blot analysis of glioma patient sera at dilution 1:50, 1:100, 1:500 revealed the presence of the 90 kDa fragment of VE-cadherin (sVE). (D) Deglycosylation Assay of sVE in serum shows, using two different antibodies to sVE that the soluble fragment is glycosylated.