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Assessement of rapamycin effect on MCF-7 breast cancer cell line and FRAP1 gene by flurescent in situ hybridization.
Egypt (IFMSA-Egypt) - Assiut University, Assiut
South Egypt Cancer Institute
Prof. Dr. Moustafa Elsharkawy
Dr. Walaa Talaat Kamel Raafat
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
Breast cancer is the world's most common cancer among women and most common cause of their death. MCF-7 is the most studied human breast cancer cell line in the world, and results from this line have had a fundamental impact upon breast cancer research and patient outcomes. Mammalian target of rapamycin (mTOR) is a crucial mediator of tumor of progression and may be a promising target in a significant proportion of patients with breast cancer. As an immunosuppressive drug, rapamycin was approved by (USA food and drug administration) in 1991 for prevention of renal allograft rejection.subsequent studies shows that rapamycin can also act as a cytostatic agent slowing or arresting growth of cell lines derived from different tumor types as breast cancer. More ANGPTL7 gene at human chromosome IP36.22 located within intron 28 of FRAP1 gene.FRAP1 phosphrylates P70 (S6K) on Thr-389,a residue whose phsphorylation is rapamyvin sensitive in vivo and necessary for S6 kinase activity,also FKBP12- rapamycin associated protein regulates translation initiation and entry into cell cycle.
What is the aim of the project?
1-detection of rapamycin effect on breast cancer cell line (MCF-7) by assessment of cell growth and viability. 2-detection of the effect of rapamycin on FRAP1 gene by flourescent in situ hybridization (FISH).
What techniques and methods are used?
1- MCF-7 cells will be used, maintained in phenol red free DMEM/F-12 medium supplemented with 10% FBS (fata bovine serum) and 1% antibiotic will be divided into 2 groups : A)control group, MCF-7 cell lines to which rapamycin will not be added. B) treatment group to MCF-7 to which rapamycin will be added at different concentrations. 2- the cell count and viability will be assessed either by trapan blue dye method or 3-(4,5 dimetyylthiazol-2- yl)-2.5 diphenyltetrazolium bromide (MTT) dye reduction (method on treatment group to determine effect of rapamycin on it .) 3- Detection of FRAP1 gene by 1p36 FISH probe.
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student will actively participate in MCF-7 cells grouping, detection of FRAP1 gene , data analysis and results comprehension are expected to be done under supervision of the professor first, then done by student's own, and the professor will review and discuss the results with them at the end.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Lectures concerning the topic and presentation of the clinical skills used.
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 scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
It's prefered if the student has studied oncology.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students
- Dilloing M.B; Dias P.; Shaprio D.N; Germanian G.S; Johanson R.K and Houghton P.J (1994): rapamycin selectively inhibits the growth of the childhood rhabdomyosarcoma cells through inhibition of signaling via the type I insulin-like growth factor receptor. Cancer Res. 1996;54:903-907
- Dumont F.J and Su Q. (1996): mechanism of actio of immunosuppresant rapamycin. Life Sci. 1996;58:373-395
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