Effect of extracellular matrix in prostatic smooth muscle cells in vitro

Author(s)

Daniel Andrés Osorio Rodríguez

• Document type: Doctoral Thesis
• Press: Campinas, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
• Defense date: 2019-09-20

Examining board members

Hernandes Faustino de Carvalho; Manoel Francisco Biancardi; Cristina Pontes Vicente; Sérgio Luis Felisbino; Luis Antonio Justulin Junior

• Advisor: Hernandes Faustino de Carvalho; Aline Mara dos Santos

Abstract

The prostate is an accessory gland of the male reproductive tract and is responsible for the production and storage of prostate fluid. This gland is affected by numerous diseases such as prostatitis, hyperplasia and cancer. These diseases compromise men¿s health and well being and are responsible for a large number of deaths, worsening with the advanced age. At histological level three compartments can be identified: epithelium, lumen and stroma. Smooth muscle cells (SMCs) are the main component of the stroma and showing a phenotypic modulation depending on different factors, one of them being the extracellular matrix (ECM). In this work we evaluated the effect of different components of ECM (collagen, fibrin and Matrigel) on adhesion, morphology, phenotype and focal adhesion kinase (FAK) localization in prostate SMCs. Preliminary results led us to study cell adhesion in collagen and subsequent blockade of Rho GTPases proteins. Finally, we study the effect of TGF'alfa' and TGF'beta' on cell migration. The results show that collagen promote a fast adhesion of SMCs, whereas, fibrin is degraded by the cells and that on Matrigel they formed cell clusters. In collagen, the actin cytoskeleton organizes into large stress fibers, which are not observed in fibrin or Matrigel. FAK had a nuclear localization in the cells seeded on collagen. On fibrin and Matrigel nuclear labeling was absent and its distribution in the cytoplasm was in as aggregates. The phosphorylated form of FAK on tyrosine 925 shows a greater co-location with the myosin heavy chain (MYH11) on collagen. SMCs on fibrin and Matrigel decreased markers of contractile phenotype, suggesting that these components of ECM favor the synthetic phenotype. The effects of inhibition of Rho GTPases proteins on the adhesion of CMLs on collagen, Rac1, Rac3 and Cdc42 are important for cell adhesion, while ROCK inhibition did not inhibit adhesion of SMCs to collagen and subsequently allowed migration - an event that did not happen when SMCs were only on collagen. Exposure to ROCK inhibitor triggered changes in cell morphology, actin cytoskeleton, ?-actinin distribution, and decreased expression of SM-actin and MYH11. Finally, we observed that TGF? stimulates the migration of CMLs, even at low concentrations. The results presented here demonstrate that ECM influences the behavior and phenotype of prostate CMLs. Rac and Cdc42 GTPases are important for cell adhesion while ROCK has an effect on the maintenance of contractile phenotype (AU)

• FAPESP's process: 15/03235-4 - Prostatic smooth muscle cells: regulation of differentiation and activation of focal adhesion kinase (FAK)by interactions with the epithelium and the extracelluar matrix
• Grantee: Daniel Andrés Osorio Rodríguez
• Support Opportunities: Scholarships in Brazil - Doctorate