| Full text | |
| Author(s): |
Nascimento, Camila F.
[1]
;
Gama-de-Souza, Leticia N.
[1, 2]
;
Freitas, Vanessa M.
[1]
;
Jaeger, Ruy G.
[1]
Total Authors: 4
|
| Affiliation: | [1] Univ Sao Paulo, Dept Cell & Dev Biol, Inst Biomed Sci, Sao Paulo - Brazil
[2] UFES, Dept Morphol, Hlth Sci Ctr, CCS, Vitoria, Espirito Santo - Spain
Total Affiliations: 2
|
| Document type: | Journal article |
| Source: | MICROSCOPY RESEARCH AND TECHNIQUE; v. 73, n. 2, p. 99-108, FEB 2010. |
| Web of Science Citations: | 12 |
| Abstract | |
Migration, invasion and protease activity are essential for tumor progression and metastasis. Metastatic cells rely on invadopodia to degrade and invade extracellular matrix (ECM). Invadopodia are membrane protrusions with enzymes required for ECM degradation. These protrusions contain cortactin and membrane type I matrix metalloproteinase (MT1-MMP) superimposed to areas of digested matrix. Here we characterized invadopodia in a cell line (CAC2) derived from human adenoid cystic carcinoma. We carried out fluorescent-substrate degradation assay to assess in situ protease activity of CAC2 cells. Digestion spots in fluorescent substrate appear as black areas in green background. Cells were cultured on Matrigel-gelatin-FITC and fixed after 1 h and 3 h. CAC2 cells were double labeled to actin and cortactin. Cells were also double stained to actin and MT1-MMR Samples were studied by laser scanning confocal microscopy. In all time points CAC2 cells showed actin, cortactin, and MT1-MMP colocalized with digestion spots in fluorescent substrate. We searched for other proteases involved in invadopodia activity. We have previously demonstrated that MMP9 influences adenoid cystic carcinoma behavior. This prompted us to investigate role played by MMP9 on invadopodia formation. CAC2 cells had MMP9 silenced by siRNA. After I h in fluorescent substrate, cells with silenced MMP9 showed clear decrease in matrix digestion compared with controls. No differences were found in cells with silenced MMP9 grown for 3 h on fluorescent substrate. Our results showed that CAC2 cells exhibit functional invadopodia containing cortactin and MT1-MMR Furthermore, MMP9 would be required in the initial steps of invadopodia formation. Microsc. Res. Tech. 73:99-108, 2010. (C) 2009 Wiley-Liss, Inc. (AU) | |
| FAPESP's process: | 07/05447-2 - Protease activity of podosomes in a cell line derived from human adenoid cystic carcinoma |
| Grantee: | Camila Nascimento Mantelli |
| Support Opportunities: | Scholarships in Brazil - Scientific Initiation |