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Telomere length and immortality of tumor cells
Austria (AMSA) - University of Vienna, Vienna
Department of Medicine I, Division of Cancer Research,
Univ.Prof.Dr. Herbert Watzke
Univ.Prof.Dr. Klaus Holzmann
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
Most cells of the adult body have limited ability to proliferate and to growth infinite in vitro. The reason is intrinsic to these cells by the absence of any mechanisms for telomere maintenance. Telomeres are the protective caps located at the ends of chromosomes that become shorter with each cell division, and after a limited number the proliferation stops and cellular ageing (senescence) is triggered. In contrast, tumour cells demonstrate activation of a telomere maintenance mechanism (TMM) to gain unlimited proliferation ability, one of the essential properties and hallmarks known in cancer biology.
What is the aim of the project?
The aim of this laboratory exercise is to determine the telomere length in tumor cells that use different TMMs.
What techniques and methods are used?
Basic cell culture, isolation of DNA from cells and tissue, quantitative real-time PCR for relative telomeric content determination, statistics, data presentation.
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?
In the lab, students will be familiarized with in vitro cultivation of tumor cells and the production of biological materials such as DNA. The relative telomere length is measured by quantitative real-time PCR (qPCR) device after pipetting probes into 96-well plates. The efficiency of PCR is determined by means of standard dilution series and used for the exact calculation of cycle threshold (Ct) values. Students must perform an accurate laboratory protocol to ensure confirmability. All analyzes are carried out as multiple measurement from which the accuracy of the data is determined. The Ct raw data of qPCR will be exported from the device software and arranged tabulated in an appropriate program. Relative quantities (RQ) values of the measured telomere repeats are calculated normalized to results of single-copy gene segments and compared to studied samples and cells with the delta-delta-Ct method. The calculated relative telomere lengths are presented in graphs and descriptive statistical analysis. The plausibility of the data, results and possible deviations between the analyzed samples are discussed and summarized in a preliminary written draft. Students will discuss and present their data and progress at weekly meetings with other members of the research group.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
First will be a discussion about appropriate experimental designs and the creation of an implementation plan for the experiments. Advantages and disadvantages of selected analytical methods should be presented in a first seminar after searching and reading of recommended literature.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a presentation - The student will prepare a scientific report - The student’s name will be mentioned in a future publication
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Basic theoretical knowledge about the research topics, but no technical skills are needed. Course is recommended for advanced medical students or post graduate students. Students are very welcome with some preliminary practical experience in cell or molecular biology background and lab work.
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) - Students in biomedical fields
- Cawthon. Telomere measurement by quantitative PCR. Nucleic Acids Res. 2002 May 15;30(10):e47.
- Lau et al. Detection of alternative lengthening of telomeres by telomere quantitative PCR. Nucleic Acids Res. 2013 Jan;41(2):e34.
- Sampl et al. Expression of telomeres in astrocytoma WHO grade 2 to 4: TERRA level correlates with telomere length; telomerase activity; and advanced clinical grade. Transl Oncol. 2012 Feb;5(1):56-65.
- Cawthon. Telomere length measurement by a novel monochrome multiplex quantitative PCR method. Nucleic Acids Res. 2009 Feb;37(3):e21.
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