Validacija novih ciljnih gena transkripcijskog faktora GLI3 u staničnim linijama tumora glave i vrata čovjeka

master's thesis

• Title (english): Validation of new target genes of transcription factor GLI3 in human head and neck tumor cell lines
• Author: Nikolina Pastva
• Mentor: Vesna Musani (mentor)
• Mentor: Inga Urlić (komentor)
• Committee member: Inga Urlić (predsjednik povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Committee member: Romana Gračan (član povjerenstva)
• Committee member: Domagoj Đikić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2024-02-01, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Tumori glave i vrata (HNSCC) su je šesti najčešći dijagnosticirani tumor u svijetu. Pacijenti dijagnosticirani s HSNCC imaju lošiju prognozu i limitirane opcije tretmana. Terapije koje ciljaju specifične mete u tim tumorima su još uvijek u razvoju, zato je nužno provesti istraživanja koja su usmjerena na bolje razumijevanje biologije HNSCC, kao i identifikaciju novih biomarkera koji će omogućiti ranije otkrivanje bolesti, ali i predstavljati nove mete za ciljane terapije. Mnoga istraživanja potvrđuju da je signalni put Hedgehog-GLI (HH-GLI) jedan od glavnih čimbenika nužnih za razvoj i održavanje HNSCC. Signalni put HH-GLI je inače uključen u procese embriogeneze, održavanja populacije matičnih stanica i regeneracije tkiva. Abnormalna aktivacija signalnog puta HH-GLI je povezna s nastanokm i razvojem brojnih tumora. Glavni akteri signalnog puta HH-GLI, transkripcijski faktori GLI1, GLI2 i GLI3 potiču prekomjernu ekspresiju proonokogenih gena od kojih je velik broj neidentificiran. Identifikacija i validacija ciljnih gena će omogućiti bolje razumijevanje biologije HNSCC. U tu svrhu sam provela dva nezavisna eksperimenta na dvije stanične linije HNSCC. Stanice su u jednom eksperimentu bile tretirane s inhibitorima signalnog puta HH-GLI, a u drugom s plazmidnim vektorima koji kodiraju GLI3 konstrukte. Analizom rezultata kvantitativnog real-time PCR-a potvrđeno je 6 gena koji su regulirani transkripcijskim faktorom GLI3, što ih čini atraktivnim metama za nove terapijske pristupe.

Abstract (english)

Tumors of the head and neck (HNSCC) are the sixth most diagnosed tumors in the world. Patients diagnosed with HNSCC have a poor prognosis and limited treatment options. Therapies targeting specific targets within the tumor are still in development, which is why it is necessary to conduct research focused on a better understanding of the biology of HNSCC, as well as the identification of new biomarkers for early disease prediction and as new targets for targeted therapies. Many studies confirm that the Hedgehog-GLI (HH-GLI) signaling pathway is one of the main factors necessary for the development and maintenance of HNSCC. The HH-GLI signaling pathway is otherwise involved in embryogenesis, maintenance of stem cell, and tissue regeneration. Abnormal activation of the HH-GLI signaling pathway is associated with the tumorigenesis of numerous tumors. This is enabled via transcription factors GLI1, GLI2, and GLI3 who promote the overexpression of pro-oncogenic genes, many of which remain unidentified. The identification and validation of these genes will enable a better understanding of the biology of HNSCC. For this purpose, I conducted two independent experiments on two HNSCC cell lines. In one experiment, cells were treated with HH-GLI signaling pathway inhibitors, and in the other, they were transfected with plasmid vectors encoding GLI3 constructs. Analysis of the quantitative real-time PCR results confirmed the regulation of 6 genes by the transcription factor GLI3, making them prime targets for new therapeutic approaches.

• Keywords: signalni put Hedgehog-GLI; GLI3; HNSCC; litij klorid; forskolin; qRT-PCR
• Language: croatian
• URN:NBN: urn:nbn:hr:217:658618
• Study programme: Title: Experimental Biology-University graduate programme; Study programme type: university; Study level: graduate; Academic / professional title: sveučilišni/a magistar/magistra eksperimentalne biologije (sveučilišni/a magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2026-02-01
• Created on: 2024-02-27 11:26:48