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Gene promoter’s methylation in the diagnosis and prediction of patients with prostate cancer
Spain (IFMSA Spain) - University of Granada, Granada
Dpt. of Biochemistry and Molecular Biology III and Immunology, Faculty of Medicine, Avda. de la Investigación 11, Granada, 18016, Spain
Jesús M. Torres de Pinedo
Jesús M. Torres de Pinedo, Sara Jordá Climent
Type of Research Project
- Basic science
What is the background of the project?
Prostate Cancer (PCa) is one of the most commonly diagnosed tumors among men in developed countries. The prostate gland depends on androgen stimulation for its development and growth. However testosterone (T) is not the major androgen responsible for growth of the prostate. T is converted to dihydrotestosterone (DHT) by the enzyme 5alpha-Reductase (5a-R) in prostatic epithelial and stromal cells. To date, the only currently used biomarker for detecting and monitoring the efficacy of treatments for PCa is the serum levels of prostate specific antigen (PSA). DNA methylation is an important mechanism for gene expression regulation, and plays an essential role in the initiation and progression of tumors.
What is the aim of the project?
The main objective of this study is to quantify in tumor tissue samples with different malignancy degrees, the methylation pattern of CpG islands in the promoter regions of the genes involved in T metabolism in the prostate
What techniques and methods are used?
Extraction of DNA from formaldehyde treated samples of tumoral prostate tissue. Treatment of amplified DNA with a bisulfate conversion kit. This treatment converts cytosine nitrogenous bases in uracil nitrogenous bases, however it is not capable of converting methylated cytosine bases. Amplification of DNA segments using polymerase chain reaction. During this amplification the methylation is lost, but the cytosine bases now indicate the previously methylated nucelotides. Methylation Sensitive-High Resolution Melting (MS-HRM) analysis on the DNA samples. This consists in the denaturalization of the DNA through the application of heat. The temperature at which the double stranded DNA becomes single stranded DNA indicates the base composition (uracil:cytosine ratio) and therefore the degree of methylation.
What is the role of the student?
- The tasks of the student will be performed on his/her own
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
- Read the bibliography related with the project provided in the project form - Get to know the basic functioning of a molecular biology laboratory - Practise using basic apparatus such automatic micropipettes, centrifuges, gas extraction cabins, thermal cycler for real-time polymerase chain reaction (PCR), DNA extraction from formalin-fixed paraffin embedded tissue (FFPE-tissue), DNA electrophoresis in agarose gel, DNA quantification by spectrophotometry - At the start the student will watch, then carry out the techniques under supervision, and then, hopefully, independently.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The tutor will teach the student about the theory behind the project and the apparatus used while carrying out the processes in the laboratory, but not separate theory lessons are given.
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?
They should know the theoretical basic of PCR and epigenetic regulation and expression.
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 - Students in biomedical fields - Dental medicine students (IADS members)
- Cancer Res 2004; 64:5956-5962 CpG hypermethylation of MDR1 gene contributes to the pathogenesis and progression of human prostate cancer. Enokida H1; Shiina H; Igawa M; Ogishima T; Kawakami T; Bassett WW; Anast JW; Li LC; Urakami S; Terashima M; Verma M; Kawahara M; Nakagawa M; Kane CJ; Carroll PR; Dahiya R.
- Cancer Lett. 2014; 342(2):248-56; Epigenetic biomarkers in prostate cancer: Current and future uses. Chiam K1; Ricciardelli C; Bianco-Miotto T
- N Engl J Med 2008; 358:1148-59; Epigenetics in cancer. Esteller M1.
- Eur Urol 60 (2011); 753-766; Epigenetics in prostate cancer: biologic and clinical relevance. Jerónimo C1; Bastian PJ; Bjartell A; Carbone GM; Catto JW; Clark SJ; Henrique R; Nelson WG; Shariat SF.
- Science (2001) Aug 10; 293; 1068-70; The role of DNA methylation in mammalian epigenetics. Jones PA1; Takai D.
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