Uloga proteina NME1 i NME2 u proliferaciji i aktivnosti signalnih puteva stanične linije melanoma čovjeka WM793B

master's thesis

• Title (english): The role of proteins NME1 and NME2 in proliferation and activity of signaling pathways in WM793B melanoma cell line
• Author: Marko Dužević
• Mentor: Ignacija Vlašić (mentor)
• Mentor: Gordana Maravić Vlahoviček (mentor)
• Committee member: Gordana Maravić Vlahoviček (predsjednik povjerenstva)
• Committee member: Sanja Dabelić (član povjerenstva)
• Committee member: Neda Slade (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of biochemistry and molecular biology); Zagreb
• Defense date and country: 2022-02-25, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Melanom je najagresivniji tip tumora kože koji najčešće nastaje zbog hiperaktivacije signalnog puta
MAPK kao posljedice onkogenih mutacija u nekoj od komponenti toga puta. U oko 60% slučajeva
mutacija BRAF p.V600E uzrokuje konstitutivno aktivnu kinazu za koju je razvijen specifičan i
efikasan inhibitor vemurafenib. Međutim, u velikom broju pacijenata javlja se otpornost na ovaj lijek
već nekoliko mjeseci nakon početka liječenja. Cilj ovog diplomskog rada bio je istražiti ulogu
nukleozid-difosfatnih kinazi NME1 i NME2 u regulaciji glavnih signalnih puteva i njihov utjecaj na
proliferaciju u svrhu ispitivanja njihovog terapijskog potencijala. Koristeći metode Western blot
analize i testa MTT utvrđena je smanjena ekspresija proteina NME u stanicama podlinija otpornih na
vemurafenib za koje je uočeno da slabije proliferiraju u odnosu na parentalni soj stanica. Također,
utišavanje gena NME komplementarnim siRNA nije utjecalo na proliferaciju stanica, no smanjilo je
ekspresiju biljega proliferacije i negativno utjecalo na aktivnost signalnog puta PI3K/AKT. Dobiveni
rezultati daju zanimljiv temelj za detaljnija istraživanja usmjerena na koji način proteini NME
reguliraju signalni put PI3K/AKT kao i otkrivanje njihovog terapijskog potencijala u liječenju
melanoma čovjeka.

Abstract (english)

Melanoma is the most aggressive type of skin tumor that most often occurs due to hyperactivation of
the MAPK signaling pathway as a consequence of oncogenic mutations in one of the components of
that pathway. In about 60% of cases the presence of BRAF p.V600E mutation results in a constitutively
active kinase for which a specific and effective inhibitor, vemurafenib, has been developed. However,
several months from the beginning of the treatment the resistance to drug occurs in many melanoma
patients. This thesis aimed to investigate the role of nucleoside diphosphate kinases NME1 and NME2
in the regulation of main signaling pathways and their impact on proliferation, with the goal to test their
therapeutic potential. Using Western blot analysis and MTT assays we observed reduced expression of
NME proteins in vemurafenib-resistant sublines that were shown to have reduced proliferation rate
compared to the parental line. In addition, NME silencing with complementary siRNA had no effect
on the cell proliferation, however it reduced the expression of a proliferation marker and negatively
influenced the activity of the PI3K/AKT signaling pathway. The obtained results provide an interesting
basis for more detailed research that is focused on how NME proteins regulate the PI3K/AKT pathway
as well as on their therapeutic potential in melanoma treatment.

• Keywords: NME1/2; melanom; vemurafenib; signalni putevi; proliferacija
• Keywords (english): NME1/2; melanoma; vemurafenib; signaling pathways; proliferation
• Language: croatian
• URN:NBN: urn:nbn:hr:163:560384
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-03-14 14:35:19