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| Author(s): |
Maíra de Assis Lima
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) |
| Defense date: | 2020-02-04 |
| Examining board members: |
Vanessa Morais Freitas;
Alexandre Bruni Cardoso;
Ana Iochabel Soares Moretti;
Marinilce Fagundes dos Santos
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| Advisor: | Vanessa Morais Freitas |
| Abstract | |
The progression of cancer depends not only on the abilities acquired by neoplastic cells, but also on the interaction between cells and their microenvironment. Extracellular matrix (ECM) components are involved in various aspects of tumor biology, providing not only solid support for cells, but also cytokines and growth factors. ADAMTSs (disintegrin and metalloproteinase with thrombospondin motifs ) are secreted proteases dependent on Zn 2+ / Ca2+, are involved in several processes and are part of the ECM. In a previous project, we demonstrated that progesterone influences on the gene and protein levels of ADAMTSs in ovarian cancer and it has been shown to reduce the migratory and invasive capacity of NIH-OVCAR-3 and ES-2 cells when compared to control cells. Our goal was to investigate by which mechanism progesterone leads to decreased invasion and migration of CHO, NIH-OVCAR-3 and ES-2 and whether the protease studied is involved in this process. For this, ovary-derived cells had ADAMTS-1 levels modulated, either with increase or decrease, and were compared with parental cells. In addition, cells with the modulated ADAMTS-1 gene and parental cells underwent progesterone treatment and were compared to untreated cells. From these groups it was observed that treatment with 500 nM or 1 µ M progesterone decreased cell migration and invasion, within 24 hours or less, without affecting viability, and the use of the progesterone receptor antagonist, RU486, recovered the migratory and invasive capacity. Both treatments, with the medium enriched with ADAMTS-1 or progesterone, led to a decrease in cell migration and invasion when compared to controls. On the other hand, the decresed levels of ADAMTS-1 does not seem to prevent the decrease of cellular migration when the cells are treated with progesterone. By Immunoblot, it was observed that treatment with 1 µ M progesterone decrease the phosphorylated form of Src and FAK, molecules that participate in the signaling cascade involved in the activation of cell migration and invasion. Using the GLISA ® kit evaluating the activity of Rho GTPases, we measured the active GTP form of small G proteins from cell lysates. The Cdc42-GTP signal increased in the decresed levels of ADAMTS-1 and tended to decrease in cells treated with ADAMTS-1-enriched medium. These data were confirmed by analysis of the transfected Cdc42 biosensor in ES-2 cells, which demonstrated that this GTPase is more active and present in the migration edge of the cells with decresed levels of ADAMTS-1 compared to the wild type cells. Therefore, we conclude that ADAMTS-1 and progesterone inhibit proliferation, polarization, migration and growth without adhesion of ovarian cancer cells. This effect has been shown to be independently to the hormone and protease studied, but does not rule out that there may be a joint work between these molecules in reducing pro-tumorigenic effects. (AU) | |
| FAPESP's process: | 15/19773-5 - Functions of progesterone and ADAMTS 1 protease in the migration of cells derived from ovarian cancer |
| Grantee: | Maíra de Assis Lima |
| Support Opportunities: | Scholarships in Brazil - Doctorate |