Određivanje obrasca lncRNA molekula PACER i ZEB1-AS1 u pacijenata s kalcificirajućom stenozom aortnih zalistaka srca

master's thesis

• Title (english): Profile of PACER and ZEB1-AS1 long non-coding RNAs in patients with calcific aortic valve stenosis
• Author: Nikola Pavlović
• Mentor: Frane Paić (mentor)
• Mentor: Vesna Benković (sumentor)
• Committee member: Maja Matulić (predsjednik povjerenstva)
• Committee member: Renata Šoštarić (član povjerenstva)
• Committee member: Jasna Lajtner (član povjerenstva)
• Committee member: Petra Peharec Štefanić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2020-12-14, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Novije spoznaje ukazuju na izuzetno važnu regulatornu ulogu dugih nekodirajućih RNA (lncRNA) molekula u intrauterinom i postnatalnom razvoju i homeostazi krvožilnog sustava. No unatoč tome, izuzetno malo se zna o ekspresijskom obrascu i funkciji molekula lncRNA u razvoju i progresiji kalcificirajuće stenoze aortnih zalistaka srca (CAVS). Cilj ovog diplomskog rada bio je odrediti ekspresijski obrazac lncRNA PACER i ZEB1-AS1 u perifernoj cirkulaciji pacijenata s kalcifirajućom stenozom aortnih zalistaka srca kao i zdravih kontrolnih ispitanika te odrediti povezanost ekspresije cirkulirajućih lncRNA PACER i ZEB1-AS1 s učestalošću kalcifirajuće stenoze aortnih zalistaka srca. Kvaliteta i kvantiteta ukupne RNA izolirane uporabom modificiranog reagensa Trizol iz uzoraka krvne plazme pacijenata s kalcifirajućom stenozom aortnih zalistaka srca i kontrolnih ispitanika odredila se pomoću spektrofotometra NanoDrop. Nakon reverzne transkripcije, analiza ekspresije ciljanih molekula lncRNA PACER i ZEB1-AS1 izvršila se metodom kvantitativne lančane reakcije polimerazom u stvarnom vremenu, uporabom CFX-96 qRT-PCR detekcijskog sustava. Statističkom analizom je utvrđeno da je ekspresija lncRNA PACER u perifernim uzorcima krvi pacijenata s kalcificirajućom stenozom aortnih zalistaka srca značajno veća u odnosu na ekspresijske vrijednosti detektirane u perifernoj krvi kontrolnih ispitanika, a lncRNA ZEB1-AS1 u perifernim uzorcima krvi pacijenata s kalcificirajućom stenozom aortnih zalistaka srca i kontrolnih ispitanika nije zabilježena.

Abstract (english)

Recent studies suggest an extremely important regulatory role of long noncoding RNA (lncRNA) molecules in intrauterine and postnatal development and vascular homeostasis. Nevertheless, extremely little is known about the expression profile and function of lncRNA molecules in the development and progression of calcific aortic valve stenosis (CAVS). The aim of this thesis was to determine the expression profile of lncRNA PACER and ZEB1-AS1 in the peripheral circulation of CAVS patients as well as in healthy control subjects and compare the quantity of circulating cell-free lncRNA PACER and ZEB1-AS1 expression with the frequency of calcification. Total RNA was isolated using modified Trizol reagent from blood plasma samples of CAVS patients and control subjects, the quality and quantity of RNA was determined using a NanoDrop spectrophotometer. After reverse transcription, the expression analysis of the target lncRNA molecules PACER and ZEB1-AS1 was performed by quantitative PCR method. Statistical analysis showed that the expression of lncRNA PACER in peripheral blood samples of CAVS patients was significantly higher than the expression values detected in the peripheral blood of control subjects, and the expression of lncRNA ZEB1-AS1 in peripheral blood samples of CAVS patients and control subjects was not detected.

• Keywords: nekodirajuće RNA; aortna stenoza; ZEB1-AS1; PACER; qRT-PCR
• Keywords (english): lncRNAs; aortic stenosis; PACER; ZEB1-AS1; qRT-PCR
• Language: croatian
• URN:NBN: urn:nbn:hr:217:147057
• 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: 2021-01-28 10:44:47