Optimizacija metode multipleks lančane reakcije polimerazom za umnažanje mikrosatelitnog biljega gena Dishevelled 3 u meningeomima čovjeka

master's thesis

• Title (english): Optimization of multiplex polymerase chain reaction method for the amplification of microsatellite marker of Dishevelled 3 gene in human meningiomas
• Author: Josip Skoko
• Mentor: Nives Pećina-Šlaus (mentor)
• Mentor: Nada Oršolić (mentor)
• Committee member: Nada Oršolić (predsjednik povjerenstva)
• Committee member: Petar Kružić (član povjerenstva)
• Committee member: Dubravko Pavoković (član povjerenstva)
• Committee member: Duje Lisičić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2015-02-13, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Meningeomi su spororastudi, benigni, primarni intrakranijalni tumori koji nastaju iz mekih moždanih ovojnica, preciznije iz meningotelijalnih stanica arahnoideje. Prema Svjetskoj zdravstvenoj organizaciji (SZO) klasificiraju se kao benigni (I. stupnja), atipični (II. stupnja) i anaplastični ili maligni (III. stupnja). Važnu ulogu pri embrionalnom i neuralnom razvoju, staničnoj proliferaciji i diferencijaciji različitih organizama ima signalni put Wnt koji je uključen i u proces tumorigeneze. Aktivacija puta odvija se preko membranskih receptora Frizzled, a može ga aktivirati oko 20 različitih liganda Wnt-a. Prvi citoplazmatski protein koji sudjeluje u prijenosu signala je fosfoprotein Dishevelled 3. Ukoliko je signalni put Wnt aktivan dolazi do inaktivacije degradacijskog kompleksa što rezultira nakupljanjem beta-katenina u citoplazmi, a zatim i u jezgri i to dovodi do transkripcije ciljanih gena i u konačnici do procesa tumorigeneze. Multipleks lančana reakcija polimerazom metoda je koja nam omogućuje analizu genetičkih promjena i genomske nestabilnosti gena DVL3. Metodu smo uspješno optimizirali i specifično umnožili miskrosatelitni biljeg gena DVL3 kao i kontrolne biljege SHGC-68373 i APEX1. Mikrosatelitni biljeg pokazao se uspješnim u otkrivanju 2 MSi (9,52%) i 5 amplifikacija (23,81%) gena DVL3 u našem uzorku meningeoma. Daljnom analizom gena DVL3 utvrditi demo njegovu ulogu u tumorigenezi meningeoma

Abstract (english)

Meningiomas are frequent benign tumours derived from the arachnoid cells. It has been well documented that wnt genes and other components of wnt signaling are critical for mammalian embryogenesis. Wnt signaling is essential in development because it acts as a regulator of the embryonic cell patterning, proliferation, differentiation, cell survival and apoptosis. The Wnt proteins bind to receptors of the Frizzled family on the cell surface. Through several cytoplasmic relay components, the first one being protein Dishevelled, the pathway is activated and the beta-catenin destruction complex is inhibited. The result is accumulation of beta-catenin in cytoplasm, which then enters the nucleus to activate transcription of Wnt target genes and tumorigenesis begins. Multiplex polymerase chain reaction is method used to analyze genetic changes and genomic instability of gene DVL3. We have achieved optimization of the method and we have specifically amplified the microsatellite marker of Dishevelled 3 gene along with control markers SHGC-58373 i APEX1. Further research is needed to establish the role of DVL3 gene and Wnt signaling in tumorigenesis of menigioma. The marker proved useful in revealing 2 MSI (9,52%) and 5 amplifications (23,81%) of DVL3 gene in our meningioma sample.

• Keywords: tumori; signalni put; Wnt; optimizacija
• Keywords (english): tumor; signaling pathway; WNT; optimization
• Language: croatian
• URN:NBN: urn:nbn:hr:217:540787
• Study programme: Title: Experimental Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-04-24 08:38:10