Ramadoss, Archana. Contribution of 3q26-29 gene cluster to glioma invasion. 2015, PhD Thesis, University of Basel, Faculty of Science.
Available under License CC BY-NC-ND (Attribution-NonCommercial-NoDerivatives).
Official URL: http://edoc.unibas.ch/diss/DissB_11658
We used SOX2 expressing human glioma cell lines, LN319 and U373 to test our hypothesis in vitro. Lentiviruses expressing shRNAs against PIK3CA, MFN1, OPA1 or SOX2 were used for genetic knockdown. Engineered cells were assayed for invasion and migration using Boyden chamber and wound healing assays, respectively. Chromatin immuno-precipitation and luciferase assays were used to demonstrate protein-DNA interactions and trans-activation of 3q26-29 genes by SOX2.
Our results show that cells downregulated for 3q26-29 genes exhibited enhanced invasion and migration, while shSOX2 and shPIK3CA cells exhibited reduced proliferation rates compared to sh scramble controls. Furthermore, we show that SOX2 knockdown reduced gene and protein expression of PIK3CA, MFN1 and OPA1 except for PIK3CA at the protein level. Chromatin immuno-precipitation assays suggested that SOX2 binds to the upstream region of 3q26-29 gene promoters in the glioma cells.
Preliminary luciferase assays in HEK293 cells suggested that SOX2 trans-activates PIK3CA and OPA1. Preliminary immunofluorescence analysis showed that cells knocked-down for 3q26-29 genes demonstrated altered mitochondrial morphology compared to sh scramble controls. Overall, our results show that SOX2, PIK3CA, MFN1 and OPA1 contribute to glioma invasion and that SOX2 is a potential regulator of the 3q26-29 genes.
|Advisors:||Mariani, Luigi and Rüegg, Markus A. and Christofori, Gerhard|
|Faculties and Departments:||03 Faculty of Medicine > Departement Biomedizin > Department of Biomedicine, University Hospital Basel > Brain Tumor Biology (Mariani)|
|Bibsysno:||Link to catalogue|
|Number of Pages:||1 Online-Ressource (193 Seiten)|
|Last Modified:||15 Jul 2016 07:12|
|Deposited On:||15 Jul 2016 07:09|
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