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Cell cycle regulatory protein expression in anoikis-resistant endothelial cells submitted to syndecan-4 silencing

Grant number: 19/21382-5
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): January 01, 2020
Effective date (End): December 31, 2020
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Carla Cristina Lopes de Azevedo
Grantee:Bianca Zaia Franco Ferreira
Home Institution: Instituto de Ciências Ambientais, Químicas e Farmacêuticas (ICAQF). Universidade Federal de São Paulo (UNIFESP). Campus Diadema. Diadema , SP, Brazil

Abstract

Anoikis is a type of regulated cell death that occurs due to lack of adhesion or improper adhesion of the cell to the extracellular matrix (ECM). Many tumor cells are anoikis resistant; this facilitates the invasion of these cells and contributes to tumor metastasis. Previous data from our laboratory showed that anoikis-resistant endothelial cells show morphological changes, high proliferation rate, low adhesion to fibronectin, laminin and collagen IV, cell cycle dysregulation, accompanied by increased expression of syndecan-4 (SDC4). SDC4 is a central mediator in cell adhesion, migration and proliferation and can interact directly and indirectly with other ECM molecules and cytoskeleton proteins during cancer progression and metastasis development. Despite having important roles in various cellular events, the specific role of syndecan-4 in cell cycle dysregulation in tumor cells is not yet fully elucidated. Thus, this work aims to understand the role of syndecan-4 in the cell cycle progression in anoikis-resistant endothelial cells that underwent the syndecan-4 gene silencing assay (Adh-shRNA-Syn4-EC), Anoikis-resistant parental endothelial cells (Adh-EC), wild endothelial cells (EC) and EJ-ras oncogene transfected endothelial cells (EJ-ras EC) regarding the expression of regulatory proteins expressed at different stages of the cycle, such as cyclins and some inhibitors (p21 e p27). The expression levels of these proteins will be evaluated by western blotting, real time PCR (qPCR) and flow cytometry techniques. Thus this study will bring new insights for understanding the mechanisms of cell cycle dysregulation during cancer progression, and may help in the development of new anticancer drugs and better direction in the treatment of the disease.

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