zde - 1. lékařská fakulta - Univerzita Karlova

3. MÍSTO
SEKCE PREGRADUÁLNÍ TEORETICKÁ – VÍTĚZNÉ PRÁCE
THE ROLE OF SHREW-1 GENE IN HUMAN GLIOBLASTOMA
MULTIFORME CELLS
Authors: Nikol Mládková, 5. roč.
Supervisor: David Cory Adamson, Molecular Neuro-Oncology Laboratory, Duke University
Medical Center
Objective: I performed series of experiments to monitor change in proliferation, migration and adhesion
in human glioblastoma multiforme (GBM) cell lines due to SHREW-1 gene knock-down.
Introduction: Tumors of the central nervous system are the second commonest tumors in children and the sixth commonest
in adults. GBM accounts for 30% of all primary tumors in adults. Survival is usually less than a year. Shrew-1 is a novel membrane
protein involved in adherens junctions. It is not expressed in about 20% of human glioblastoma multiforme tumors. The
suppression of the gene may change clinically relevant cell behavior.
Methods: SHREW-1 was knocked down in U-251 cell line using RNA interference and confirmed by cDNA synthesis and PCR.
Proliferation was measured using MTT Proliferation Assay in five-day time course, cell migration was measured using Wound
Assay and Transwell Migration Assay toward Growth Factor in six- and twelve-hour time course, cell adhesion was measured on
laminin coated wells.
Results: The proliferation rate of U-251 cell line was lower in SHREW-1 knock-down (KD) cells compared to control. The average
proliferation ratio of control cells compared with KD cells is 1,56× higher in five-days time course. The transwell migration
study showed 1,83× higher average ratio of cell migration after 6 hours in KD cells compared to control. In 12 hours, the average
ratio of cell migration would be 1,23× higher in KD cells compared to control. The Wound Assay showed that KD cells do not
heal within 48 hours while control cells fully healed at this time point. The adhesion to laminin of KD cells is 1,23× higher compared
to control.
Conclusions: Shrew-1 loss decreases proliferation of U-251 cells, it enhances migration in transwells, however it decreases
migration in Wound Assay, probably due to loss of cell polarity and possibly change in proliferation. It also changes the adhesion
of the cells to extracellular matrix protein – the KD cells are more prone to adhere to laminin coated wells.
Nikol Mládková obsadila 3. místo v sekci pregraduální teoretické. Děkan fakulty předává cenu a blahopřeje.
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