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Enhancing iodide-related therapeutic options in thyroid cancer
Portugal (PorMSIC) - University of Lisbon, Lisbon
Laboratório de Genética da Faculdade de Medicina da Universidade de Lisboa, AvenidaProfessor Egas Moniz, 1649-028 Lisboa – Portugal
Ana Luísa Silva
Type of Research Project
- Basic science
What is the background of the project?
The sodium iodide symporter (NIS) is highly expressed in thyroid tissue. Since the expression of NIS results in the accumulation of iodide, its expression in tumor cells allows the use of radioactive iodine (131I) as a diagnostic and therapeutic tool. It is clinical relevant to develop strategies directed at NIS expression and regulation so as to increase iodide uptake in refractory tumors enabling treatment with 131I.The induction of the p38 mitogenic kinase activity by the GTPase RAC1 has been shown to be involved in the stimulation of NIS expression. In a recent study of our group, the overexpression of an hyperactive variant of RAC1, the RAC1b protein, was shown to be associated with a subset of thyroid carcinomas with unfavorable outcome, carrying the activating B-Raf V600E mutation.
What is the aim of the project?
The aim of this project is to investigate whether RAC1/1b signaling has an impact on the modulation of NIS expression, as a potential approach to enhance the efficiency of iodide uptake.
What techniques and methods are used?
Some of the basic technics of molecular and cell biology, will be employed in this study, including: gene cloning, cell culture and transfection; RNA and protein extraction, RT-PCR (Reverse transcription polymerase chain reaction) and quantitative RT-PCR and Western blot. With all of this technique we will be able to determine if the RAC1/1b has an impact on the modulation of NIS expression, its effects and the consequences in the uptake of iodine.
What is the role of the student?
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student will be allowed to participate in the execution of most of the methodologies used in this study, under supervision of lab fellows participating in this research project. This will allow the student to learn a wide range of techniques used in a molecular biology lab, from cloning to gene expression and signaling pathway analysis. Particularly, the student will learn how to: appropriately use all the equipment regularly used in the lab, including balances, pipettes, electrophoresis and centrifuges; clone a gene into a plasmid vector (from RNA extraction, reverse transcription, polymerase chain reaction, ligation, bacterial transformation, and DNA extraction); transfect plasmids in a cell line and extract protein to assess and quantify expression using Western blotting as well as extract RNA to assess and quantify expression using qPCR.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Preliminary readings will be needed - a list of papers will be provided and a brief lecture will help the student to consolidate the gathered information.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
To understand the basics of molecular and cell biology.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Pre-Medical students from the American-British system
- Spitzweg C; Bible KC; Hofbauer LC; Morris JC. Advanced radioiodine-refractory differentiated thyroid cancer: the sodium iodide symporter and other emerging therapeutic targets. Lancet Diabetes Endocrinol. 2014 Oct;2(10):830-42. doi:10.1016/S2213-8587(14)70051-8.
- Kogai T; Liu YY; Mody K; Shamsian DV; Brent GA. Regulation of sodium iodide symporter gene expression by Rac1/p38β mitogen-activated protein kinase signalling pathway in MCF-7 breast cancer cells. J Biol Chem. 2012 Jan 27;287(5):3292-300.doi: 10.1074/jbc.M111.315523.
- Silva AL; Carmo F; Bugalho MJ. RAC1b overexpression in papillary thyroid carcinoma: a role to unravel. Eur J Endocrinol. 2013 Apr 29;168(6):795-804. doi:10.1530/EJE-12-0960.
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