Gene expression of membrane transporters ABCB1, ABCG2, SLC22A1 and SLCO1A2 in cell lines treated with commercial inhibitor of JAK-STAT pathway

• Grant number: 12/01081-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): June 01, 2012
• Effective date (End): December 31, 2012
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Elvira Maria Guerra Shinohara
• Grantee: Guilherme Wataru Gomes
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Myelofibrosis (MF) is a Myeloproliferative Neoplasm (MPN) Philadelphia negative characterized by fibrosis development in bone marrow, in response to cell proliferation. Discover of JAK2V617F mutation and its relation with MF made possible the development of drugs targeted to the inhibition of JAK2 protein. However, little information is known about the mechanisms involved in inhibitors of JAK-STAT pathway cellular influx and efflux, which may represent resistance mechanisms and response variance to the drug. This study aims to examine the effect of a commercial inhibitor of JAK-STAT pathway in regulating the expression of genes that encode membrane transporters ABCB1, ABCG2, SLC22A1, and SLCO1A2, and evaluate their expression of them in different cell lines. HEL921.7, SET2 (positive for JAK2V617F), HepG2, and Caco-2 cell lines will be cultured and treated with different commercial inhibitors of JAK-STAT pathway concentrations for different periods. After checking for growth and cell viability, mRNA will be extracted from the cells and cDNA synthesized to evaluate the expression of genes ABCB1, ABCG2, SLC22A1, and SLCO1A2 in real-time RT-PCR. The expression of membrane transporters encoded by these genes will be assessed by flow cytometry. With the results of this study, we hope to contribute to a better understanding of the pharmacokinetics of inhibitors of JAK-STAT pathway and possible mechanisms involved in drug response variability.(AU)