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To investigate the role of PSMB8 in invasion and angiogenesis in human glioblastoma cells
Taiwan (FMS-Taiwan) - National Defense Medicine Center, Taipei
Department of Biology and Anatomy
Associate Professor Ying Chen
Type of Research Project
- Basic science
What is the background of the project?
Glioblastomas, one of the most common brain tumors, have not developed efficient therapies currently. A proteasome component proteasome subunit beta type-8 (PSMB8), reveals as a necessary survival gene in glioblastoma cells. Recent study demonstrates that PSMB8 inhibition reduces glioblastoma cells proliferation and migration.Invasion and angiogenesis are also important for glioblastoma cells progression. However, the role of PSMB8 in cell invasion and vascular angiogenesis is unknown. Therefore, small interferenceribonucleic acid (RNA) andinducible knockdown of PSMB8 will be used to evaluate the role of PSMB8 in glioblastoma cell invasion and angiogenesis. Furthermore, in vivo study will be performed to prove the hypothesis in vitro in the future.
What is the aim of the project?
The aim of this project is to identify the role of PSMB8 in angiogenesis in human glioblastoma. We hope that PSMB8 is a candidate gene in glioblastoma therapy.
What techniques and methods are used?
Glioblastoma cells (LN229 and U87MG) are transfected with PSMB8 interference RNA (siRNA) or short hairpin RNA (shRNA). The RNA and protein expression of PSMB8 will be evaluated by real time polymerase chain reaction (RT-PCR) and Western blotting. Then, the conditioned medium of glioblastoma cells will be collected and incubated with endothelial cells. The tube formation, migration and invasion of endothelial cells will be studied to check the ability of angiogenesis. In conclusion, these results will indicate whether PSMB8 inhibition in human glioblastoma reduce the angiogenetic ability of endothelial cells.
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?
The human glioblastoma cells and endothelial cells will be cultured. PSMB8 siRNA or shRNA will be transfected to the cell. After transfection, the RNA and protein expression of PSMB8 will be evaluated by real time-PCR and Western blotting. After check whether the knockdown are successful or not,the culture medium will collected by the student as conditioned medium. The following steps will be taken also by the student. The endothelial cells will be cultured with the conditioned medium and tested for the ability of angiogenesis by tube formation assay, migration and invasion. Then, the results will be photographed by the camera and analyzed by ImageJ software.Finally, these results let us known the role of PSMB8 inhibition of glioma cells in endothelial cells angiogenesis.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The angiogenesis-related paper reading and presenting in our lab meeting.
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 presentation - The student will prepare a scientific report - The student will prepare an abstract - 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?
subject passed: biology and chemistry
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students
- Interference with PSMB4 Expression Exerts an Anti-Tumor Effect by Decreasing the Invasion and Proliferation of Human Glioblastoma Cells.Cell Physiol Biochem. 2018;45(2):819-831. Cheng Y.-C. • Tsai W.-C. • Sung Y.-C. • Chang H.-H. • Chen Y. https://www.karger.com/Article/FullText/487174
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