Utjecaj prekomjerne ekspresije proteina SH2D3C na regulaciju signalnog puta NRF2 - KEAP1

master's thesis

• Title (english): The impact of overexpression of SH2D3C protein on the regulation of NRF2 - KEAP1 signaling pathway
• Author: Ivana Marinović
• Mentor: Mihaela Matovina (mentor)
• Mentor: Nada Oršolić (komentor)
• Committee member: Nada Oršolić (predsjednik povjerenstva)
• Committee member: Zorana Grubić (član povjerenstva)
• Committee member: Jasna Lajtner (član povjerenstva)
• Committee member: Domagoj Đikić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2022-12-01, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Analizom interaktoma humane dipeptidil-peptidaze III (DPP III) otkriven je protein 3C koji
sadrţi domenu SH2 (SH2D3C) kao jedan od potencijalnih interaktora DPP III, proteina koji je
uključen u regulaciju oksidacijskog stresa preko signalnog puta NRF2-KEAP1. Potencijalna
uloga interakcije DPP III-SH2D3C još uvijek je nepoznata, ali poznato je da protein SH2D3C
sudjeluje u regulaciji stanične migracije i adhezije, organizacije tkiva i regulaciji imunog
odgovora te bi interakcija DPP III-SH2D3C mogla predstavljati vezu izmeĎu regulacije tih
procesa i oksidacijskog stresa. Cilj ovog rada bio je prekomjerno eksprimirati izoforme 2 i 3
proteina SH2D3C u stanicama HEK293T te utvrditi kako povećana ekspresija ovih izoformi
djeluje na regulaciju signalnog puta NRF2-KEAP1, odnosno na ekspresiju mRNA gena NQO1,
HMOX1 i NRF2 u bazalnim i u uvjetima oksidacijskog stresa izazvanog vodikovim peroksidom.
Prema dobivenim rezultatima prekomjerna ekspresija izoforme 2 u bazalnim uvjetima ne utječe
na ekspresiju gena NQO1, HMOX1 i NRF2, dok prekomjerna ekspresija izoforme 3 uzrokuje
statistički značajno smanjenje ekspresije gena NQO1 i NRF2. U uvjetima oksidacijskog stresa
nakon tretmana vodikovim peroksidom prekomjerna ekspresija obiju izoformi ne utječe na
ekspresiju gena NQO1, HMOX1 i NRF2.

Abstract (english)

Analysis of the human dipeptidyl peptidase III (DPP III) interactome revealed SH2 domaincontaining protein 3C (SH2D3C) as one of the potential interactors of DPP III, a protein
involved in the regulation of oxidative stress via the NRF2-KEAP1 signaling pathway. The
potential role of the DPP III-SH2D3C interaction is still unknown, but it is known that the
SH2D3C protein participates in the regulation of cell migration and adhesion, tissue organization
and the regulation of the immune response, and DPP III-SH2D3C interaction could represent a
link between the regulation of these processes and oxidative stress. The aim of this thesis was to
overexpress isoforms 2 and 3 of the SH2D3C protein in HEK293T cells and to determine how
the increased expression of these isoforms affects the regulation of the NRF2-KEAP1 signaling
pathway, i.e. the mRNA expression of the NQO1, HMOX1 and NRF2 genes in basal and in
conditions of oxidative stress caused by hydrogen peroxide. According to the obtained results,
the overexpression of isoform 2 in basal conditions does not affect the expression of NQO1,
HMOX1 and NRF2 genes, while the overexpression of isoform 3 causes a statistically significant
decrease in the expression of the NQO1 and NRF2 genes. In conditions of oxidative stress after
treatment with hydrogen peroxide, overexpression of both isoforms does not affect the
expression of NQO1, HMOX1 and NRF2 genes

• Keywords: DPP III; SH2D3C; NRF-KEAP1; kvantitativni PCR
• Keywords (english): DPP III; SH2D3C; NRF-KEAP1; qPCR
• Language: croatian
• URN:NBN: urn:nbn:hr:217:583502
• 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; Embargo expiration date: 2023-12-01
• Created on: 2022-12-20 13:02:55