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Name of the research project (Highly specific and matching the aim of the project) The biological roles of miRNAs in cisplatin resistance of WW domain containing oxidoreductase-negative breast cancer cells
Turkey (TurkMSIC) - Namik Kemal University, Tekirdag
Department of Medical Biology, Tekirdag Namik Kemal University School of Medicine, Namik Kemal Campus Street Number: 1,Tekirdag, 59030, Turkey
PhD Bahadir Batar
PhD Bahadir Batar
Type of Research Project
- Basic science
What is the background of the project?
Please, provide the background of the project (Provide information about the project; min. 10 lines, max. 25 lines) WW domain-containing oxidoreductase (WWOX) is a tumor suppressor gene spanning the fragile site FRA16D. The WWOX gene encodes a 46-kDa molecular weight protein containing two N-terminal WW domains responsible for protein-protein interactions and a short-chain dehydrogenase (SDR) domain that plays a role in steroid metabolism. Wwox protein is reduced or absent in about two thirds of breast cancer cases. Wwox protein loss is a frequent event in triple-negative breast cancer (TNBC). We first showed that Wwox loss or reduction significantly contributes to cisplatin resistance of TNBC cells and provides a survival advantage to cancer cells. However, the underlying molecular mechanism of Wwox-deficient TNBC cells resistance to cisplatin remains elusive. MicroRNAs (miRNAs) are small short non-coding RNA molecules about 22 nucleotides long. miRNAs play an emerging role in essential biological activities, such as the development of resistance to targeted chemotherapeutic agents and ionizing radiation (IR). MEMEDER (Breast Health Association) is an associaton in Turkey that has been established in 2007 and does various awareness actions about breast cancer. According to the data of this association, every year in Turkey we diagnose 25.000 women for breast cancer. There is also a foundation named MEVA (Turkish Breast Foundation) that does similiar actions with MEMEDER.
What is the aim of the project?
The aim of this research project is to identify the biological roles of miRNA’s in cisplatin resistance of Wwox-deficient breast cancer cells.
What techniques and methods are used?
What techniques and methods are used? Mention the steps/stages in this project. (Provide examples and avoid abbreviations; which tools will be used in general in this project?) Cell culture: Cell culture is the process by which cells are grown under controlled conditions, generally outside their natural environment. After the cells of interest have been isolated from living tissue, they can subsequently be maintained under carefully controlled conditions. We will use the cell culture majorly for the TNBC cells. Bacterial transformation: Bacterial transformation is a process of horizontal gene transfer by which some bacteria take up foreign genetic material (naked DNA) from the environment. We will use bacterial transformation to transfer the WWOX gene. transient transfection: When cells are transiently transfected with plasmids, the DNA is introduced into the nucleus of the cell, but does not integrate into the chromosome. This means that many copies of the gene of interest are present, leading to high levels of expressed protein. Western blot: Western blot is a method used to detect specific protein molecules from among a mixture of proteins. Western blots can also be used to evaluate the size of a protein of interest, and to measure the amount of protein expression. Cell cycle analysis with the usage of Flow cytometry/FACS (Fluorescent Activated Cell Sorter); Cell cycle analysis by DNA content measurement is a method that most frequently employs flow cytometry to distinguish cells in different phases of the cell cycle. Cell cycle anomalies revealed on the DNA content frequency histogram are often observed after different types of cell damage, for example such DNA damage that interrupts the cell cycle progression at certain checkpoints. Such an arrest of the cell cycle progression can lead either to an effective DNA repair, which may prevent transformation of normal into a cancer cell (carcinogenesis), or to cell death, often by the mode of apoptosis. apoptosis assay: apoptosis assay detects and quantifies the cellular events associated with programmed cell death, including caspase activation, cell surface exposure of phosphatidylserine (PS) and DNA fragmentation.
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
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
What are the tasks expected to be accomplished by the student? (Minimum 10 lines, provide detailed explanation) The following tasks are expected to be accomplished by the student: -Learning the cell culture steps and process such as preparing the cell culture broth, sub-culturing and splitting. -Analysing the cell lines by using Flow cytometry/FACS. Understanding the difference between a cancer cell line and non-cancer cell line. -Investigating the steps and process of western blot and afterwards trying to western blot by the students her/his self with the help of the supervisor. -Data analysis of the project process/results with the usage of SPSS 26 (Statistical Package for Social Sciences) and MS Excel (with supervision if necessary).
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Will there be any theoretical teaching provided for the student (preliminary readings, lectures, courses, seminars)? Please describe how much teaching and by whom. Theoretical teaching in the form of presentations will be given by PhD Bahadir Batar at least once per week depending on the availibility of the teacher. The subjects: “The cell cycle in cancer cells, cell death mechanisms, general inforamation about breast cancer, the laboratory methods that will be 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 poster - 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?
No spesific skills are required
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)
- Schrock MS; Batar B; Lee J; Druck T; Ferguson B; Cho JH; Akakpo K; Hagrass H; Heerema NA; Xia F; Parvin JD; Aldaz CM; Huebner K. Wwox-Brca1 interaction: role in DNA repair pathway choice. Oncogene. 2017; 36: 2215-2227.
- Schrock MS; Karras JR; Guggenbiller MJ; Druck T; Batar B; Huebner K. Fhit and Wwox loss-associated genome instability: A genome caretaker one-two punch. Adv Biol Regul 2017; 63: 167-176.
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