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Cardoso ACF. Effects of shear stress exposure on melanoma cells to tumor vascular remodeling [thesis]. São Paulo: “Faculdade de Medicina, Universidade de São Paulo”; 2018. Melanoma pathogenesis and staging are clinically well characterized, although a large variation is observed in progression and therapeutic responses. Intrinsic mechanisms of melanoma cells contribute to this phenomenon, such as phenotypic plasticity and the expression of certain molecules as galectin-3. Galectin-3 plays a prominent role in tumor growth, metastasis, and to build a functional vascular network. In this way, melanoma has alternative mechanisms of vascularization, in which both endothelial cells and tumor cells form the luminal surface of vessels and are in contact with flowing blood. Here, we postulated that shear stress promotes cell adaptation and melanoma cells communicate with (1) other tumor cells in order to support tumor progression; or (2) with endothelial cells to ensure vascular maintenance. We compared two methods for generating shear stress, namely revolution of concentric plates and stirring of a suspended cone on a plate, which is the gold standard method for studies on endothelial cells. The concentric plates model yielded more reliable results, since it ensured the greater viability and adhesion of cells up to 24h, as compared to the cone and plate system. We then investigated the angiogenic potential of melanoma cells by monitoring the activation of endothelial cells upon exposure to the conditioned medium (CM) from cells exposed to shear stress. Shear stress significantly increased the secretion of Angiopoietin-2, IL-8 and PLGF, in a galectin-3 independent manner. Also, shear stress decreased the growth rate of galectin-3 positive melanoma cells, but not when galectin-3 was down regulated. No such effect was observed with the shearderived CM. Instead, we did observed long-term melanoma survival under low density upon treatment with the shear-derived CM. In this scenario, the expression of galectin-3 did not change the results. Regarding the effect of shear-derived CM to vascular remodeling, endothelial cells did not show any change in cell proliferation. However, the CM of melanoma cells upon shear accelerated the endothelial cell tube formation and disorganized the actin stress fibers. CM also decreased the phosphorylation of ERK. Taken together, these data indicate that the production of proangiogenic and proinflammatory molecules by melanoma cells under shear enables long-term survival and proliferation of naïve melanoma cells. To endothelial cells, shear-derived CM led to changes in ERK signaling and cytoskeletal rearrangement. Descriptors: melanoma; galectin 3; shear strength; cell plasticity; angiogenesis modulating agents; vascular remodeling.

Consequências da exposição de células de melanoma ao shear stress para o remodelamento da microvasculatura intratumoral