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Etablierung und Validierung der RNA-Interferenzmethode am Beispiel Apoptose-relevanter Gene

dc.contributor.advisorKugler, Wilfried PD Dr.de
dc.contributor.authorKöhler, Franziskade
dc.date.accessioned2013-01-14T15:27:12Zde
dc.date.available2013-01-30T23:50:53Zde
dc.date.issued2012-07-05de
dc.identifier.urihttp://hdl.handle.net/11858/00-1735-0000-000D-EFDD-2de
dc.identifier.urihttp://dx.doi.org/10.53846/goediss-1566
dc.format.mimetypeapplication/pdfde
dc.language.isogerde
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/de
dc.titleEtablierung und Validierung der RNA-Interferenzmethode am Beispiel Apoptose-relevanter Genede
dc.typedoctoralThesisde
dc.title.translatedEstablishment and validation of RNA interference using the example of apoptosis-relevant genesde
dc.contributor.refereeLakomek, Max Prof. Dr.de
dc.date.examination2012-07-24de
dc.subject.dnb610 Medizin, Gesundheitde
dc.subject.gokMED 000de
dc.description.abstractengSpecific inhibition of gene expression is becoming an increasingly important method for experimental strategies to influence tumor growth on a molecular level. In this work, we investigated the role of individual molecules for erucylphosphocholin (ErPC) induced apoptosis using specific, synthesized small interfering RNAs (siRNAs). The human gliobastoma cell lines A172, U118MG and U373MG were used as model system; protein expression was measured with western blot analysis. As a first step, we confirmed that the RNA interference method is in fact applicable to our in vitro model system for human brain tumor cells using a simple, morphological test. In the second step, we optimzied the RNA interference method for our model cell lines by targeting the mRNA of the cytoskeleton protein Lamin A/C. The transfection parameters optimized were the cell count and the concentrations of siRNA and transfection reagent Oligogectamin. The specificity of the employed method was confirmed using a control siRNA targeting Luciferase. In the third step, we applied the RNA interference method employing our optimized parameters to the genes APAF-1, Caspase 2 and Caspase 3, which are known to participate in cell apoptosis. We were able to show that the expression of the respective proteins can be down regulated in different glioblastoma cell lines using specific siRNA. The last part of this work investigates the functional importance of Caspase 3 and APAF-1 “knock downde
dc.contributor.coRefereeBastians, Holger Prof. Dr.de
dc.contributor.thirdRefereeHanisch, Uwe-Karsten Prof. Dr.de
dc.subject.topicMedicinede
dc.subject.gerRNA-Interferenzde
dc.subject.gerApoptosede
dc.subject.gerErucylphosphocholinde
dc.subject.gerCaspase 3de
dc.subject.gerCaspase 2de
dc.subject.gerApaf-1de
dc.subject.gerGlioblastomzellende
dc.subject.engsiRNAde
dc.subject.engapoptosisde
dc.subject.engerucylphosphocholinede
dc.subject.engcaspase-3de
dc.subject.engCaspase-2de
dc.subject.engapaf-1de
dc.subject.enggliomade
dc.subject.bk44.00de
dc.identifier.urnurn:nbn:de:gbv:7-webdoc-3602-3de
dc.identifier.purlwebdoc-3602de
dc.affiliation.instituteMedizinische Fakultätde
dc.identifier.ppn728919729de


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