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Study of sphingosine kinase 1 and 2 proteins in the response to hypoxia and reoxygenation in an oral keratinocyte lineage model

Grant number: 22/14844-5
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): December 01, 2022
Effective date (End): November 30, 2023
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Andréia Machado Leopoldino
Grantee:Laura Stefany Zamara
Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:16/19103-2 - SET and sphingolipids in head and neck squamous cell carcinoma: signaling, targets and antiumoral therapy, AP.TEM

Abstract

Sphingosine-1-phosphate (S1P), a bioactive sphingolipid, is produced by sphingosine kinase 1 (SK1) and SK2 and acts through paracrine, autocrine and intracellular signaling pathways. Changes in S1P levels, SK1 and SK2 enzymes, as well as S1P receptors, have been described in cancer. Furthermore, S1P have also been recognized as important cellular mediator in hypoxia and angiogenesis. Hypoxia is a key feature of solid tumors associated with a worse prognosis and greater aggressiveness. Although the role of SK1 and S1P has been extensively studied, the involvement of SK2 in hypoxia and reoxygenation is not well understood. In this project, we will investigate the role of SK1 and SK2 in the response to hypoxia and reoxygenation in an oral keratinocyte (NOK) lineage model with and without SK2 and SK1 overexpression compared to the control cell (NOK with vector empty). After exposing the cells to hypoxia and reoxygenation conditions, metabolic parameters such as glucose uptake, lactate release, and cell survival, as well as angiogenesis stimulation will be analyzed. We will also analyze the protein levels of the main signaling pathways involved in the processes studied by Western blot and/or immunofluorescence and angiogenesis through the tubule formation assay using human umbilical vein endothelial cells (HUVEC). The results will contribute to the characterization of the role of SK2 compared to SK1 in hypoxia and reoxygenation and, consequently, to define which sphingosine kinase has greater relevance for the processes studied and for the development of oral cancer.

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