Opredelitev vloge izbranih krožnih RNA pri raku jeter

Jug, Oto (author), Režen, Tadeja (mentor)

Krožne RNA so nov tip molekul RNA, ki jim njihova struktura omogoča večjo stabilnost. Namen magistrske naloge je bil preučiti njihov vpliv na lastnosti celic. Glede na transkriptomske študije smo izbrali krožne RNA, ki so diferencialno izražene v hepatocelularnem karcinomu; hsa_circ_0001806, hsa_circ_0004913 in hsa_circ_0036044. Meritve smo izvajali na celičnih linijah Huh-7 in Hep-G2, ki so uporabljene kot celični modeli za analize v povezavi s hepatocelularnim karcinomom. Celice smo transfecirali s krožnimi RNA in nato merili vpliv na metabolno aktivnost celic z uporabo AlamarBlueTM in reagenta CCK-8. Pri meritvah s prvim reagentom je biološka variabilnost presegala učinek krožne RNA na celice. Pri uporabi reagenta CCK-8 pa smo uspeli pridobiti rezultate z manj variabilnosti. Merili smo tudi vpliv krožnih RNA na hitrost migracije celic s preizkusom hitrosti zapiranja raze, kar smo spremljali s slikanjem pod svetlobnim mikroskopom. Uspešno smo optimizirali metodo transfekcije, merjenja metabolne aktivnosti ter merjenja hitrosti zapiranja raze. Vendar nismo izmerili statistično značilnih učinkov krožne RNA na metabolizem ali hitrost migracije celic.

• Language: Slovenian
• Keywords: Medicinska biotehnologija, rak jeter, krožna RNA
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [O. Jug]
• Year: 2020
• PID: 20.500.12556/RUL-117794
• UDC: 606:616.36-006.6:601.4:577.21(043.2)
• COBISS.SI-ID: 26811395
• Publication date in RUL: 26.07.2020

Secondary language

• Language: English
• Title: Characterisation of the role of selected circular RNA in liver cancer

Abstract

Circular RNA are a recently discovered type of RNA molecule, defined by their increased lifespan. Purpose of this thesis was to discern their influence on cells. We chose circular RNAs that were expressed differently in hepatocellular carcinoma based on transcriptomic studies. We selected: hsa_circ_0001806, hsa_circ_0004913, and hsa_circ_0036044. Measurements were made on Huh-7 and Hep-G2 cell lines, which are used as cellular models for analysis concerning hepatocellular carcinoma. We measured metabolic cell activity with AlamarBlueTM and CCK-8 after cells were transfected with circular RNA. The use of the first reagent proved inadequate as biological variability surpassed the effects of circular RNA on the cells. The second reagent, CCK-8, provided results with less variability. We also measured the effects of circular RNA on cell migration using wound healing assays, which we measured with a light microscope. We were successful at optimising the transfection method as well as measuring the metabolic activity and cell migration, but we did not measure any statistically significant influence of circular RNA on cellular metabolic activity or cell migration.

• Keywords: Medical biotechnology, hepatocellular carcinoma, circular RNA