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Expressionshemmung von MCM3 in Glioblastomen durch RNA-Interferenz

dc.contributor.advisorLakomek, Max Prof. Dr.
dc.contributor.authorLauerer, Peter
dc.titleExpressionshemmung von MCM3 in Glioblastomen durch RNA-Interferenzde
dc.title.translatedExpression inhibition of MCM3 in glioblastomas by RNA interferencede
dc.contributor.refereeLakomek, Max Prof. Dr.
dc.description.abstractengThis work shows effects of specific suppression of MCM3-protein, which is upregulated in human glioblastoma cells (U87MG), by using RNAi. The effects on metabolic activity, cell viability, proliferation, cell cycle and apoptosis were investigated. It could be shown that specific MCM3-siRNAs can exert an inhibitory effect on the proliferation of human glioblastoma cells. Five MCM3-siRNAs, which all showed a suppression of MCM3 at the protein level were tested. Further analyzes were continued with the two most effective MCM3-siRNAs (MCM3-siRNA 13 and MCM3-siRNA 15). MCM3-siRNA 13 showed at the protein level in Western blot analyzes at a low concentration of 10 nM a MCM3-suppression of >90%, MCM3-siRNA 15 at least an inhibition of 84%. Functional analyzes about metabolic activity and proliferation as MTT assay, BrdU assay and trypan blue staining demonstrated a reduced metabolic activity and a proliferation stop of the treated U87MG cells. This was reflected in a G0 / G1 and G2 / M arrest of treated cells by FACS analyzes with propidium iodide. Notes for induction of apoptosis of U87MG cells by treatment with MCM3-siRNAs did not arise. The most effective of the two MCM3-siRNAs in Western blot as well as in the functional assays was MCM3-siRNA 13. MCM3-siRNA 13 demonstrated the highest suppression rate at protein level as well as the lowest metabolic activity and proliferation in the functional assays. In the real-time RT-PCR MCM3-siRNA 13 also showed a high suppression of MCM3 mRNA, but it was less effective than MCM3-siRNA 15. MCM3-siRNA 15 had the highest suppression of mRNA for MCM3. There are explanations why the MCM3-siRNA 13 was less effective than MCM3-siRNA 15 on the mRNA level, but this needs to be analyzed in further experiments. With a potential therapeutic siRNA treatment the proliferation of glioblastoma cells could stop or slow down. That would mean a slower expansion of the brain tumor. For patients, this would be not a cure, but it could lead to a prolonged survival with a higher quality of
dc.contributor.coRefereeBastians, Holger Prof. Dr.
dc.contributor.thirdRefereeWulf, Gerald Prof. Dr.
dc.affiliation.instituteMedizinische Fakultätde
dc.subject.gokfullOnkologie (PPN619875895)de

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