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Meningeomi su najčešći primarni tumori središnjeg živčanog sustava, koji u manje od 20% slučajeva mogu pokazivati agresivnost, recidive i metastaze. Među signalnim putevima uključenima u progresiju meningeoma nalazi se i signalni put Wnt čiji je središnji medijator protein DVL1 koji inhibira djelovanje destrukcijskog kompleksa za razgradnju ß-katenina. Uslijed toga se ß-katenin nakuplja u citoplazmi i premješta u jezgru gdje potiče transkripciju onkogena. Budući da je dokazana uloga proteina iz porodice DVL u progresiji tumora središnjeg živčanog sustava, cilj je ovog rada bio istražiti utjecaj promjena gena DVL1 na progresiju intrakranijalnih meningeoma čovjeka. Izolirana je DNA tumora i periferne krvi pacijenata te su amplificirani mikrosatelitni biljeg D1S468 i genska regija koja kodira domenu PDZ, važnu u prijenosu signala Wnt. Gel-elektroforezom umnoženog biljega D1S468 utvrđeno je postojanje mikrosatelitne nestabilnosti u 9,09% uzoraka i gubitka heterozigotnosti u 6,06% uzoraka. Analizom taljenja visoke rezolucije i Sangerovim sekvenciranjem genske regije PDZ, utvrđene su mutacije u 100% sekvenciranih uzoraka. Rezultati analize pokazali su kako dio otkrivenih mutacija ima štetan učinak na biološku funkciju proteina DVL1, mijenjanjem mjesta prekrajanja ili promjenama okvira čitanja sekvence. Ukupni rezultati ovog istraživanja pokazuju ulogu promjena gena DVL1 u razvoju intrakranijalnih meningeomima. Meningiomas are the most common primary tumours of the central nervous system, which in less than 20% of cases may show aggression, recurrence, and metastasis. Among the signalling pathways involved in meningioma progression is the Wnt signalling pathway. The central mediator of the Wnt signalling pathway is the DVL1 protein which inhibits the destruction complex that targets ß-catenin. This results in the accumulation of ß-catenin in the cytoplasm and its translocation to the nucleus, where it promotes oncogene transcription. Since it has been shown that proteins from the DVL family play a role in the progression of central nervous system tumours, the aim of this study was to investigate the influence of alterations in the DVL1 gene on the progression of human intracranial meningiomas. The microsatellite marker D1S468 and the gene region encoding the PDZ domain, important in transducing the Wnt signal, were amplified from DNA that has been isolated from tumour and peripheral blood samples of the patients. Gel electrophoresis of the amplified marker D1S468 revealed the existence of microsatellite instability in 9.09% of samples, loss of heterozygosity in 6.06% of samples. High-resolution melting analysis and Sanger sequencing of the PDZ gene region revealed mutations in 100% of sequenced samples. The results of the analysis showed that some of the detected mutations have a deleterious effect on the biological function of the DVL1 protein, by altering the splice site or shifting the reading frame of the sequence. The overall results of this study demonstrate the role of alterations in DVL1 gene in intracranial meningiomas |
