Optimized zymogram protocol from 3D spheroid cultures to study MMP-2 and -9 activities in tumor cells.

ABSTRACT: Three-dimensional spheroids are more representative of tumors than cell-cultured monolayers. As in tumors, gradients of oxygen, nutrients and wastes are found in spheroid cultures but not in classical cultured monolayers. On the other hand, cell-based assays on the latter are hardly applicable to spheroid cultures. Such is the case for zymogram assays, which are classically used to measure MMP-2 and MMP-9 activities, and for immunoblots to measure the phosphorylation of proteins involved in ligand-induced intracellular signaling in normal and tumor cells. In this study we used two renal cancer cell lines as models, the first derived from a pediatric rhabdoid tumor and the second from an adult clear cell renal cell carcinoma. Using these two cell lines, we successfully developed a simple in

SUBMITTER: Majo S 

PROVIDER: S-EPMC11987344 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature