doktorska disertacija
Kristijan Skok (Author), Iztok Takač (Mentor), Uroš Maver (Co-mentor), Rajko Kavalar (Co-mentor)

Abstract

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.

Keywords

trojno negativni rak dojk;MFUM-BrTNBC-1;rak dojk;in-vitro modeli;celične linije;

Data

Language: Slovenian
Year of publishing:
Typology: 2.08 - Doctoral Dissertation
Organization: UM MF - Faculty of Medicine
Publisher: K. Skok]
UDC: 576/577:602.3:618.19-006+616(043.3)
COBISS: 131347459 Link will open in a new window
Views: 23
Downloads: 1
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Other data

Secondary language: English
Secondary title: Analysis of phenotypical, molecular, and genetic properties of a novel triple-negative breast cancer cell line
Secondary abstract: 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.
Secondary keywords: triple negative breast cancer;MFUM-BrTNBC-1;breast cancer;in vitro models;cell lines;
Type (COBISS): Dissertation
Pages: IX, 105 f.
ID: 14934555