Vloga NONO in PCBP2 v izražanju izbranih circRNA

Musina, Valeriya (author), Režen, Tadeja (mentor)

Krožne RNA so nov razred RNA, ki imajo kovalentno povezano verigo. Krožne RNA imajo številne funkcije, lahko delujejo kot onkogeni ali tumor supresorji in imajo spremenjeno izražanje v številnih rakih, tudi v hepatocelularnem karcinomu. Krožne RNA se sintetizirajo v procesu povratnega spajanja, ki je odvisen od kanoničnih izrezovalno-povezovalnih kompleksov in cis in trans-delujočih elementov, kot so RNA-vezavni proteini. Povišano izražanje RNA-vezavnih proteinov NONO in PCBP2 statistično značilno negativno korelira s slabšim preživetjem bolnikov z rakom jeter. Namen diplomske naloge je bil preučevanje vloge NONO in PCBP2 v uravnavanju izražanja izbranih krožnih RNA, ki imajo spremenjeno izražanje v tumorjih raka jeter. In vitro analizo smo izvedli v humani hepatocelularni celični liniji Huh-7. V okviru diplomske naloge smo uspešno prekomerno izrazili oba RNA-vezavna proteina, NONO in PCBP2, s pomočjo qPCR izmerili izražanje krožnih RNA v celicah s prekomerno izraženima proteinoma in v kontrolnih celicah ter določili relativno spremembo v izražanju izbranih krožnih RNA. Potrdili smo, da lahko RNA-vezavni proteini vplivajo na izražanje izbranih krožnih RNA.

• Language: Slovenian
• Keywords: krožne RNA, NONO, PCBP2, hepatocelularni karcinom
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2021
• PID: 20.500.12556/RUL-129977
• COBISS.SI-ID: 82932739
• Publication date in RUL: 09.09.2021

Secondary language

• Language: English
• Title: The role of NONO and PCBP2 in the expression of selected circRNAs

Abstract

Circular RNAs are a new class of RNA which have a covalently closed structure. Circular RNAs have many functions, they can act as oncogenes or tumor suppressors, and are often dysregulated in various cancers, including hepatocellular carcinoma. Circular RNAs are synthesized during the process called backsplicing that depends on canonical spliceosomal machinery and both cis-acting and trans-acting factors such as RNA-binding proteins. Upregulated expression of RNA-binding proteins NONO and PCBP2 is statistically significantly negatively correlated with poorer survival of liver cancer patients. The aim of the thesis was to examine the role of NONO and PCBP2 in the regulation of the expression of selected circular RNAs that are dysregulated in liver cancer tumors. In vitro analysis was performed in the human hepatocellular cell line Huh-7. In this thesis we successfully overexpressed both RNA-binding proteins, NONO and PCBP2, measured the expression of circular RNAs in cells with overexpressed proteins and control cells using qPCR, and determined the relative change in the expression of selected circular RNAs. We confirmed that RNA-binding proteins can affect the expression of selected circular RNAs.

• Keywords: circular RNA, NONO, PCBP2, hepatocellular carcinoma