20.500.12556/RUL-173479 Detekcija bakrovega transporterja ATP7A in ceruloplazmina v celičnih linijah in kliničnih vzorcih za napoved kemorezistence pri bolnicah z rakom jajčnikov Detection of copper transporter ATP7A and ceruloplasmin in cell lines and clinical samples for chemoresistance prediction in patients with ovarian cancer V kliničnih vzorcih jajčnikov, odvzetih med diagnostično laparoskopijo 16 bolnic z rakom jajčnikov, smo z metodo qPCR preučevali razmerje izražanja gena ATP7A za bakrov transporter ATP7A ter izražanja referenčnega gena ACTB. V krvni plazmi odvzeti pred operacijo in ascitesu teh istih bolnic smo s testom ELISA izmerili koncentracije ceruloplazmina ter nato iskali povezavo med izražanjem ATP7A/ACTB in koncentracijo ceruloplazmina v obeh tekočinah, saj sta tako ATP7A kot tudi ceruloplazmin vpletena v pridobivanje kemorezistence na citotoksična zdravila na osnovi platine (Pt-BM) pri raku jajčnikov. Povezavo med izražanjem ATP7A/ACTB in koncentracijo ceruloplazmina smo uspeli statistično značilno dokazati v krvni plazmi pred operacijo pri bolnicah z vsemi histologijami, ki niso imele znakov nekroze na jajčniku (r= 0,591), še močnejšo povezavo pa smo ugotovili pri bolnicah s seroznim karcinomom visoke stopnje malignosti (HGSC), ki niso imele znakov nekroze na jajčniku (r= 0,7639) . Ugotovili smo tudi, da so imele bolnice z vsemi histologijami, ki niso imele znakov nekroze na jajčniku, statistično značilno (p-vrednost < 0,1) višje koncentracije ceruloplazmina v plazmi pred operacijo, če je bilo razmerje ATP7A/ACTB > 1. Razmerje izražanja ATP7A/ACTB ni bilo povezano s koncentracijo ceruloplazmina v ascitesu. S poskusom znotrajceličnega barvanja in pretočne citometrije ATP7A v celičnih linijah HOSEpiC, OAW28 in HT1080 smo uspeli statistično značilno (p-vrednost < 0,05) dokazati, da je v celicah celične linije OAW28, ki je bila vzgojena iz celic bolnice s kemorezistentnim rakom jajčnikov, več ATP7A transporterjev kot v celicah linije zdravega jajčnika HOSEpiC in v celicah linije fibrosarkoma HT1080, ki so po navodilih proizvajalca primarnih protiteles služile za kontrolo vezave na ATP7A. Using the qPCR we had calculated the ratio between the expression of the gene ATP7A, which encodes the copper transporter ATP7A, and the reference gene ACTB in ovarian clinical samples, which were obtained during diagnostic laparoscopy from 16 patients with ovarian cancer. In the blood plasma and ascites of the same patients, we measured ceruloplasmin concentrations using an ELISA test and later investigated the association between the ratio of ATP7A/ACTB expression and ceruloplasmin concentrations in both fluids, since both ATP7A and ceruloplasmin are involved in the development of chemoresistance to platinum-based cytotoxic drugs (Pt-BM) in ovarian cancer. We demonstrated a statistically significant association between the ratio of ATP7A/ACTB expression and ceruloplasmin concentrations in preoperative blood plasma of patients with all histological subtypes who had shown no signs of ovarian necrosis (r = 0.591) and an even stronger association in patients with high grade serous carcinoma (HGSC) who had shown no signs of ovarian necrosis (r = 0.7639). We also demonstrated that patients with all histological subtypes who had shown no signs of ovarian necrosis had statistically significantly (p-value < 0.1) higher ceruloplasmin concentrations in preoperative plasma if the ratio of ATP7A/ACTB expression was > 1. The ratio of ATP7A/ACTB expression was not associated with ceruloplasmin concentration in ascites. Using intracellular staining coupled with flow cytometry of the ATP7A transporter in HOSEpiC, OAW28, and HT1080 cell lines, we were able to demonstrate statistically significantly (p-value < 0.05) higher levels of ATP7A in OAW28 cells, which were isolated from a patient with chemoresistant ovarian cancer, compared to cells of the healthy ovarian cell line HOSEpiC and the fibrosarcoma cell line HT1080, which served as a control for primary antibody binding with ATP7A. rak jajčnikov ATP7A ceruloplazmin kemorezistenca ovarian cancer ATP7A ceruloplasmin chemoresistance true false false [H. Suchy] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-09-18 07:15:45 2025-09-18 07:15:54 2025-11-09 03:57:52 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 616-006:604.4(043.2) 264799 249551107 52014.pdf 52014.pdf 1 46C2C8C134885E077EFA2A4593DB5F37 21f3436b595f582f2150c97456608041c3691892b2cabe21cd6a4a186c61c7b7 63a53d19-944e-11f0-9328-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=217994 Biotehniška fakulteta 0 0 0