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Title:Analiza fenotipskih, molekularnih in genetskih lastnosti nove celične linije trojno negativnega raka dojk
Authors:ID Skok, Kristijan (Author)
ID Takač, Iztok (Mentor) More about this mentor... New window
ID Maver, Uroš (Comentor)
ID Kavalar, Rajko (Comentor)
Files:.pdf DOK_Skok_Kristijan_2022.pdf (6,85 MB)
MD5: 8C6F1FCCF6D11B9687A3D246593957C9
 
Language:Slovenian
Work type:Doctoral dissertation
Typology:2.08 - Doctoral Dissertation
Organization:MF - Faculty of Medicine
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
Place of publishing:Maribor
Year of publishing:2022
PID:20.500.12556/DKUM-81503 New window
COBISS.SI-ID:131347459 New window
Publication date in DKUM:29.11.2022
Views:823
Downloads:77
Metadata:XML DC-XML DC-RDF
Categories:MF
:
SKOK, Kristijan, 2022, Analiza fenotipskih, molekularnih in genetskih lastnosti nove celične linije trojno negativnega raka dojk [online]. Doctoral dissertation. Maribor. [Accessed 27 March 2025]. Retrieved from: https://dk.um.si/IzpisGradiva.php?lang=eng&id=81503
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Licences

License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:04.04.2022

Secondary language

Language:English
Title:Analysis of phenotypical, molecular, and genetic properties of a novel triple-negative breast cancer cell line
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.
Keywords:triple negative breast cancer, MFUM-BrTNBC-1, breast cancer, in vitro models, cell lines


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