20.500.12556/DKUM-82754 Izražanja skupine genov DNA-metiltransferaz (DNMT1, DNMT3A in DNMT3B), DNA-demetilaz (TET1, TET2, TET3 in TDG) ter gena RNA-metiltransferaza TRDMT1 v tumorjih bolnic z rakom dojk magistrsko delo Expression of dna methyltransferases genes DNMT1, DNMT3A and DNMT3B, DNA demethylases genes TET1, TET2, TET3 and TDG and RNA methyltransferase gene TRDMT1 in tumors from patients with breast cancer DNA-metiltransferaze DNMT1, DNMT3A in DNMT3B, DNA-demetilaze TET1, TET2, TET3 in TDG ter RNA-metiltransferaza TRDMT1 imajo pomembno vlogo pri uravnavanju metilacije genoma in genskih prepisov, s čimer uravnavajo izražanje genov. Spremenjeno izražanje omenjenih encimov povezujemo z nastankom in razvojem različnih vrst rakov, med drugim tudi raka dojk. Mehanizmi njihovega delovanja so v literaturi dobro opisani, medtem ko njihova patofiziološka vloga pri nastanku in razvoju raka dojk ni povsem znana. Namen magistrskega dela je bil raziskati izražanje DNMT1, DNMT3A, DNMT3B, TET1, TET2, TET3, TGD in TRDMT1 na parnih vzorcih zdravega in bolnega tkiva 17 bolnic z rakom dojk ter v povezavi z gradusom in izražanostjo progesteronskih receptorjev pri raku dojk. Povišano izražanje v tumorskem tkivu v primerjavi z zdravim okolnim tkivom smo ugotovili pri DNMT1 (p<0,01), DNMT3A (p<0,05), DNMT3B (p<0,01), TET2 (p<0,05) in TET3 (p<0,01), medtem kot se izražaje TET1 ni razlikovalo med zdravim in tumorskim tkivom. Izražanje TDG in TRDMT1 je bilo višje v tumorskem kot zdravem tkivu, vendar razlika ni dosegla statistične značilnosti. Za tumorsko tkivo je bilo statistično značilno višje izražanje DNMT1 (p<0,001) in DNMT3A (p<0,001) v primerjavi z DNMT3B in statistično značilno višje izražanje TDG v primerjavi s TET1 (p<0,05) in TET3 (p<0,01). Razlik v izražanji med DNMT1 in DNMT3A ter TET1, TET2 in TET3 v tumorskem tkivu nismo ugotovili. Primerjava izražanja vseh genov med seboj je v tumorskem tkivu pokazala na statistično značilno i) višje izražanje TDG v primerjavi z ostalimi proučevanimi geni, ii) višje izražanje DNMT1 v primerjavi s TET1 (p<0,01) in TRDMT1 (p<0,05), iii) nižje izražanje DNMT3B v primerjavi s TET1 (p<0,05) in TRDMT1 (p<0,05) in iv) nižje izražanje TET3 od TET1 (p<0,05) in TRDMT1 (p<0,05). Omenjenih razlik v zdravem tkivu nismo ugotovili. Izraženost progesteronskih receptorjev in gradus tumorja nista vplivala na izražanje proučevaih genov. Post hoc analiza je pokazala na statistično značilno i) višje (p<0,05) izražanje DNMT1 v bolnem tkivu pri rakih tretjega gradusa v primerjavi z izražanjem DNMT1 v zdravem tkivu pri rakih drugega gradusa, ii) višje (p<0,05) izražanje TDG pri rakih dojk drugega kot tretjega gradusa in iii) višje izražanje TDG v tumorskem tkivu v primerjavi z izražanjem TET1 (p<0,05), TET2 (p<0,05) in TET3 (p<0,05) v zdravem tkivu ter TET3 v tumorskem tkivu (p<0,05) pri rakih drugega gradusa. Naša raziskava kaže na specifično povišano izražanje DNA-metiltransferaz DNMT1, DNMT3A in DNMT3B, DNA-demetilaz TET1, TET2, TET3 in TDG in RNA-metiltransferaze TRDMT1 pri raku dojk. V nadaljevanju bo za boljše razumevanje vloge metilacije pri tumorigenezi raka dojk potrebno rezultate izražanja posameznih metiltransferaz in demetilaz povezati z globalnimi študijami metiloma. DNMT1, DNMT3A and DNMT3B DNA methyltransferases, TET1, TET2, TET3 and TDG DNA demethylases and the TRDMT1 RNA methyltransferase play an important role in regulating DNA and RNA methylation, thereby regulating gene expression. Altered expression of these enzymes has been linked to the initiation and development of various cancers, including breast cancer. Although their mechanisms of action are well described, their pathophysiological role in the development and progression of breast cancer is not entirely characterised. The aim of the master's thesis was to investigate the expression of DNMT1, DNMT3A, DNMT3B, TET1, TET2, TET3, TGD and TRDMT1 in paired tumor and normal tissue samples from 17 breast cancer patients and in relation to a cancer stage and progesterone receptor expression. In the present study, DNMT1 (p<0.01), DNMT3A (p<0.05), DNMT3B (p<0.01), TET2 (p<0.05) and TET3 (p<0.01) showed higher expression in tumor than paired normal tissue, whereas there were no differences in TET1 expression. TDG and TRDMT1 expression was higher in tumour than in normal tissue, but the difference did not reach statistical significance. Furthermore, tumor tissue exhibited higher expression of DNMT1 (p<0.001) and DNMT3A (p<0.001) compared to DNMT3B and higher expression of TDG compared to TET1 (p<0.05) and TET3 (p<0.01). There were no differences in the expression of DNMT1 and DNMT3A and of TET1, TET2 and TET3 in tumour tissue. Comparison of gene expression across all genes showed that tumour but not normal tissue exhibited i) higher expression of TDG compared to the other genes studied, ii) higher expression of DNMT1 compared to TET1 (p<0, 01) and TRDMT1 (p<0.05), iii) lower expression of DNMT3B compared to TET1 (p<0.05) and TRDMT1 (p<0.05) and iv) lower expression of TET3 than TET1 (p<0.05) and TRDMT1 (p<0.05). Progesterone receptor expression and tumour grade did not affect the expression of the genes studied. However, post hoc analysis showed i) higher (p<0.05) expression of DNMT1 in grade 3 cancers compared to DNMT1 expression in normal tissue in grade 2 cancers, ii) higher (p<0, 05) TDG expression in grade 2 than grade 3 cancers and iii) higher TDG expression in tumour tissue compared to TET1 (p<0.05), TET2 (p<0.05) and TET3 (p<0.05) expression in normal tissue and TET3 (p<0.05) expression in tumour tissue (p<0.05) in grade 2 cancers only. Our study shows a gene-specific increased expression of DNA methyltransferases (DNMT1, DNMT3A and DNMT3B), DNA demethylases (TET1, TET2, TET3 and TDG) and the RNA methyltransferase TRDMT1 in breast cancer. In the future, to better understand the role of methylation in breast cancer tumorigenesis, it will be necessary to effectively integrate the expression levels of methyltransferases and demethylases with DNA/RNA methylation petterns on a global scale. DNA-metiltransferza DNA-demetilaza TRDMT1 rak dojk RT-qPCR DNA-methyltransferase DNA-demethylase TRDMT1 breast cancer RT-qPCR true true false [N. Šadl] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2022-08-31 18:00:44 2022-09-27 12:42:05 2022-12-07 03:06:58 0000-00-00 00:00:00 2022 0 Maribor 0 Maribor 1 spletni vir (1 datoteka PDF (XI, 57 f.)) 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 575.117:618.19-006(043.2) 132485379 MAG_Sadl_Nastja_2022.pdf MAG_Sadl_Nastja_2022.pdf 1 1A7C1E9363C94E700D9D3DA78F7EF4FB 97349211010f2364c2d09e964f5780c34cc31a45fbe2cb7ed1220828e40adbe1 73a2ab66-3dcb-11ed-9af8-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=163457 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0