20.500.12556/DKUM-81503 Analiza fenotipskih, molekularnih in genetskih lastnosti nove celične linije trojno negativnega raka dojk Analysis of phenotypical, molecular, and genetic properties of a novel triple-negative breast cancer cell line Rak dojk je najpogostejši rak na svetu. Eden izmed njegovih podtipov je trojno negativni rak dojk (TNRD), za katerega je značilen agresiven potek, slabša prognoza in heterogenost. V svoji doktorski nalogi predstavim novo izolirano celično linijo TNRD, poimenovano MFUM-BrTNBC1. Linija je bila celovito karakterizirana (morfološko, fenotipsko, genotipsko). Za kontrole so služile komercialne celične linije z različnimi receptorskimi profili (MCF-7, MDA-MB-231, MDA-MB-453). Primerjalne morfološke analize so bile opravljene s pomočjo mikroskopije ter biomedicinske slikovne analize. S pomočjo imunocito/histokemičnih barvanj je bila preverjena izraženost hormonskih receptorjev (estrogen, progesteron), HER2, p53 in Ki-67 statusa. Ekspresija receptorjev je bila dodatno preverjena na transkripcijski ravni s qRT-PCR metodo. Dodatno je bilo opravljeno RNA sekvenciranje za molekularno analizo. Avtentičnost celičnih linij je bila preverjena s pomočjo STR profiliranja. MFUM-BrTNBC-1 celična linija je ohranila trojno negativni receptorski status in genetsko stabilnost do vsaj 6. pasaže. Rezultati STR analize in navzkrižna preverba z mednarodnimi bazami so dokazali avtentičnost celične linije. MFUM-BrTNBC1 se je po morfologiji celic razlikovala tudi od TNRD celične linije MDA-MB-231. Razlike od komercialne linije, predstavljajo tudi glavne prednostne lasnosti MFUM-BrTNBC-1. Te so izolacija iz primarnega tumorskega tkiva, popoln izvorni zapis, mednarodno poimenovanje, celoten in edinstven STR profil ter genetska stabilnost do vsaj 6. pasaže. Breast cancer is the most common cancer worldwide. One of its subtypes is triple-negative breast cancer (TNBC), which is aggressive, has a worse prognosis, and is heterogeneous. In this thesis, I present a newly isolated TNBC cell line MFUM-BrTNBC-1. We performed a detailed characterization of morphological, phenotypic, and genotypic characteristics. As controls served 3 commercial breast cancer cell lines with different receptor profiles (MCF-7, MDA-MB-231, MDA-MB-453). Comparative morphological analysis was done using microscopy (light, fluorescence) and biomedical image analysis. Hormone receptor (estrogen, progesterone), p53 and Ki-67 statuses were determined by immunocyto/histochemical tests. Receptor expression was validated at the transcriptional level by qRT-PCR. RNA sequencing was utilised for additional analysis of molecular properties. Authenticity was checked by STR profiling. MFUM-BrTNBC-1 maintained the primary triple-negative receptor status and was genetically stable up to the 6th passage. STR profiles were crosschecked with international databases and confirmed authenticity. Morphologically, MFUM-BrTNBC-1 differed from the commercial TNBC cell line MDA-MB-231. The key differences and simultaneously advantages between MFUM-BrTNBC-1 and the commercial cell are, isolation from a primary tumour; good growth characteristics; identical phenotype to primary tissue; complete origin records, unique identifier; complete, unique STR profile and genetic stability up to (at least) 6th passage. trojno negativni rak dojk MFUM-BrTNBC-1 rak dojk in-vitro modeli celične linije triple negative breast cancer MFUM-BrTNBC-1 breast cancer in vitro models cell lines true true false Slovenski jezik Angleški jezik Doktorsko delo/naloga 2022-04-04 22:21:35 2022-11-29 07:08:22 2022-11-30 03:07:39 0000-00-00 00:00:00 2022 0 Maribor 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 131347459 DOK_Skok_Kristijan_2022.pdf DOK_Skok_Kristijan_2022.pdf 1 8C6F1FCCF6D11B9687A3D246593957C9 c6274498e96f11ede7ba8c1ec221fd3d4e4f3df1932025b8fddf7825f194af80 57ed4ca2-68e0-11ed-a11e-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=164486 Medicinska fakulteta 0 0 0