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Mechanisms of reversal of the multidrug resistance proteins by low intensity ultrasound in glioma stem cell
China (IFMSA-China) - China Medical University, Shenyang
Type of Research Project
- Basic science
What is the background of the project?
Glioma is a malignant brain tumor accounting for 33.3% ~ 58.9% of intracranial tumors. Patients suffering from malignant glioma will only survive for less than 18 months even undergoing surgery and radiotherapy and/or chemotherapy for the multidrug resistance. More than 180,000 to 600,000 young people die of glioma each year. Searching for targets associated with drug resistance and molecular targets is the focus of the concern of scholars at home and abroad. Glioma, a highly heterogeneous tumor, shows characteristics of the stem cells, oligodendrocytes precursor, astrocytes and oligodendrocytes. Our research team has been working on reversing the multidrug resistance of glioma by low intensity ultrasound for 10 more years and obtained relevant issues including two national natural fund projects and two provincial fund projects.
What is the aim of the project?
Find the undergoing mechanisms of reversing the multidrug resistance in glioma stem cells by low frequency and intensity ultrasound.
What techniques and methods are used?
The techniques and methods include western blot, immunofluorescence staining on cell line or animal model in this research. Western blot Cells were lysed in the ice-cold Radio Immunoprecipitation Assay(RIPA) lysis buffer (Beyotime, P0013B). The protein concentration of the lysate was determined by bicinchoninic acid (BCA) kits (Beyotime, P0010S). The protein samples (20 μg per lane) from the C6 and U87 cells were separated on 10% polyacrylamide gels using Mini Protean III electrophoresis tanks (Bio-Rad, Hercules, California). After electrophoresis, the proteins were transferred to Polyvinylidene fluoride(PVDF) membranes according to standard procedures. The polyvinylidene fluoride (PVDF) membranes were blocked for 1 h with 5% nonfat dry milk in TBST(Tris-HCl buffer salt solution+tween) buffer (20 mM Tris-HCl, pH 7.4; 150 mM NaCl; 0.1% Tween-20), followed by incubating with primary antibodies an overnight at 4°C. After washing the membranes three times with TBST the blots were incubated with corresponding secondary antibodies (Proteintech, US) for 2 hrs at room temperature. Finally, specific proteins were visualized using enhanced chemiluminescence (BIORAD, USA) and detected using ImageJ software. Immunofluorescence staining We will use double immunofluorescence staining to examine the expression of proteins. Tissue sections will be rehydrated and prepared for the application as it described for immunohistochemistry. After antigen retrieval, sections will be incubated with antibody for 1 hour. Sections will be washed in phosphate buffer saline (PBS) and then will be incubated with a combination of a secondary antibodies. Sections will be examined using the fluorescence microscope Olympus BX51 (Olympus, Tokyo, Japan) and photographed with DP71 camera (Olympus, Tokyo, Japan). Data analysis and quantification will be performed. Data will be analyzed with SPSS(Statistical Product and Service Solutions) statistical software system.
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
What are the tasks expected to be accomplished by the student?
1.Rat C6 glioma cells and human U87 glioma cells were cultured in DMEM (Dulbecco's modified eagle medium) and 10% FBS at 37°C in a humidified atmosphere containing 5% CO2. 2. The cells were cultured and then harvested for in vitro experiments. Western blot analysis and Immunofluorescence staining are needed. 3.Data analysis and quantification: data will be analyzed with SPSS (statistical software system).
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
No theoretical teaching provided
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?
Sterile operation is an operation in the sterile room or ultraclean Taichung to prevent microbes from entering the human body or pollution of antimicrobial technology operation. It is very important in the various biological experiment and production practice of the basic operation. The requirement is: before operation to kill microorganisms such as bacteria and viruses in the operating space; Ensure operation space isolation with the outside world in the process of operation, to avoid microbial invasion.
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)
- Yao Lei; Zhang Zhen. The reversal of MRP1 expression induced by low-frequency and low-intensity ultrasound and curcumin mediated by VEGF in brain glioma. OncoTargets and Therapy;2019:12 3581–3593.
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