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master's thesis
miRNA kao regulator izražaja antioksidativnih enzima u mononuklearnim stanicama periferne krvi
2024. urn:nbn:hr:152:291854
Lagator, Maja
Josip Juraj Strossmayer University of Osijek
Faculty of Medicine Osijek

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Lagator, M. (2024). miRNA kao regulator izražaja antioksidativnih enzima u mononuklearnim stanicama periferne krvi (Master's thesis). Osijek: Josip Juraj Strossmayer University of Osijek, Faculty of Medicine Osijek. Retrieved from https://urn.nsk.hr/urn:nbn:hr:152:291854

Lagator, Maja. "miRNA kao regulator izražaja antioksidativnih enzima u mononuklearnim stanicama periferne krvi." Master's thesis, Josip Juraj Strossmayer University of Osijek, Faculty of Medicine Osijek, 2024. https://urn.nsk.hr/urn:nbn:hr:152:291854

Lagator, Maja. "miRNA kao regulator izražaja antioksidativnih enzima u mononuklearnim stanicama periferne krvi." Master's thesis, Josip Juraj Strossmayer University of Osijek, Faculty of Medicine Osijek, 2024. https://urn.nsk.hr/urn:nbn:hr:152:291854

Lagator, M. (2024). 'miRNA kao regulator izražaja antioksidativnih enzima u mononuklearnim stanicama periferne krvi', Master's thesis, Josip Juraj Strossmayer University of Osijek, Faculty of Medicine Osijek, accessed 19 November 2024, https://urn.nsk.hr/urn:nbn:hr:152:291854

Lagator M. miRNA kao regulator izražaja antioksidativnih enzima u mononuklearnim stanicama periferne krvi [Master's thesis]. Osijek: Josip Juraj Strossmayer University of Osijek, Faculty of Medicine Osijek; 2024 [cited 2024 November 19] Available at: https://urn.nsk.hr/urn:nbn:hr:152:291854

M. Lagator, "miRNA kao regulator izražaja antioksidativnih enzima u mononuklearnim stanicama periferne krvi", Master's thesis, Josip Juraj Strossmayer University of Osijek, Faculty of Medicine Osijek, Osijek, 2024. Available at: https://urn.nsk.hr/urn:nbn:hr:152:291854

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Metadata
TitlemiRNA kao regulator izražaja antioksidativnih enzima u mononuklearnim stanicama periferne krvi
Title (english)miRNA as a regulator of antioxidative enzymes expression in peripheral blood mononuclear cells
AuthorMaja Lagator
MentorInes Drenjančević (mentor)
Committee memberZrinka Mihaljević (predsjednik povjerenstva)
Committee memberIvana Jukić (član povjerenstva)
Committee memberInes Drenjančević (član povjerenstva)
GranterJosip Juraj Strossmayer University of Osijek
Faculty of Medicine Osijek
Osijek
Defense date and country2024-07-12, Croatia
Scientific / art field,
discipline and subdiscipline		BIOMEDICINE AND HEALTHCARE
Basic Medical Sciences
Abstract
CILJ ISTRAŽIVANJA: Primarni cilj je uhodati metodu microRNA pomoću miRNA Taqman assay-a na humanim mononukelarnim stanicama periferne krvi. Sekundarni cilj je istražiti sekvence miRNA za gene antioksidativnih enzima te istražiti njihovu učinkovitost u utišavanju ciljanog gena hsa-miR-532-3p (GPx4). NACRT STUDIJE: Presječna studija. MATERIJALI I METODE: Materijal je uzorak mononuklearnih stanica izoliranih iz periferne venske krvi zdravog, mladog ispitanika. Za izolaciju miRNA iz mononuklearnih stanica krvi koristila se metoda s kiselim gvanidin-tiocijanatom i fenol-kloroform. Za detekciju i kvantifikaciju koristio se komercijalni kit Applied Biosystems™ TaqMan™ Small RNA Assay. Izražaj miRNA u triplikatnom uzorku odredio se real time PCR-om (BioRad, CFX96 RealTime System). Rezultati su normalizirani housekeeping genima i prikazani kao preklopna promjena (2 −∆∆𝐶𝑡). Razina značajnosti postavljena je na α = 0,05. Statističke analize provedene su korištenjem Graph Pad Prism v6.01 i Microsoft Excel 2016. REZULTATI: 2 −∆∆𝐶𝑡 ciljanog gena, hsa-miR-532-3p iznosila je 0,708 odnosno 0,685. Vrijednost 2 −∆∆𝐶𝑡 manja od 1 ukazuje na downregulaciju genskog izražaja, u ovom je slučaju došlo do ~30% smanjenog genskog izražaja u odnosu na housekeeping gene. ZAKLJUČAK: Metoda miRNA Taqman assay uspješno je validirana za analizu izražaja miRNA u humanim mononuklearnim stanicama periferne krvi. miRNA ima ulogu u regulaciji genskog izražaja antioksidativnih enzima.
Abstract (english)
OBJECTIVES: The aim of this study was to validate microRNA method using miRNA Taqman assay on human mononuclear cells. Secondary aim was to investigate miRNA sequences targeting antioxidant enzymes and their ability to regulate target genes such as GPx4. STUDY DESIGN: Cross-sectional study. MATERIALS AND METHODS: Material for this study was a sample of peripheral blood mononuclear cells isolated from a healthy, young participant. miRNA was isolated with acid guanidine-thiocyanate and phenol-chloroform method. For detection and quantification commercial kit Applied Biosystems™ TaqMan™ Small RNA Assay was used. miRNA expression was determined with real time PCR (BioRad, CFX96 Real-Time System). Results were normalized with housekeeping genes and shown as fold change (2 −∆∆𝐶𝑡). Significance level was set at α = 0.05. Statistical analyzes were performed using Graph Pad Prism v6.01 and Microsoft Excel 2016. RESULTS: 2 −∆∆𝐶𝑡 of the target gene, hsa-miR-532-3p was 0.708 in relation to housekeepin gene miR-16 and 0.685 in relation to housekeeping gene miR-423-3p. A value of 2 −∆∆𝐶𝑡 less than 1 indicates a downregulation of gene expression, in this case there was ~30% reduced gene expression. CONCLUSION: miRNA Taqman assay method was successfully validated for the analysis of miRNA expression in human peripheral blood mononuclear cells. miRNA plays a role in the regulation of gene expression of antioxidant enzymes.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:152:291854
Study programmeTitle: University graduate study
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra medicinsko laboratorijske dijagnostike (magistar/magistra medicinsko laboratorijske dijagnostike)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2024-10-11 12:09:16