20.500.12556/RUL-186666 Učinek gemcitabina na imunomodulatorne lastnosti raka trebušne slinavke in njegove posledice za z granulami posredovano citotoksičnost NK-celic Effect of gemcitabine on the immunomodulatory properties of pancreatic cancer and its consequences for granule-mediated NK cell cytotoxicity Rak trebušne slinavke je ena najagresivnejših malignih bolezni, za katero sta značilna izrazito imunosupresivno tumorsko mikrookolje in slaba odzivnost na zdravljenje. Pomemben del zdravljenja še vedno ostaja gemcitabin. Ta učinkovina lahko poleg neposrednega citotoksičnega učinka na tumorske celice vpliva tudi na njihovo komunikacijo z imunskimi celicami. Namen magistrske naloge je bil preučiti, ali kondicionirani medij celic raka trebušne slinavke, predhodno tretiranih z gemcitabinom, posredno vpliva na fenotipske in citotoksične značilnosti naravnih celic ubijalk. V raziskavi smo uporabili celični liniji raka trebušne slinavke PANC-1 in CAPAN-2, ki smo ju tretirali z gemcitabinom, nato pa iz njunih izločkov pripravili kondicionirani medij. Kondicioniranemu mediju smo izpostavili celično linijo naravnih celic ubijalk NK-92 ter primarne naravne celice ubijalke, izolirane iz krvi dveh zdravih darovalcev. Izražanje molekul, vključenih v z granulami posredovano citotoksičnostjo, smo analizirali z metodo prenosa western, izražanje površinskih označevalcev aktivacije, zaviranja in adhezije pa s pretočno citometrijo. Rezultati so pokazali, da kondicionirani medij tumorskih celic vpliva na citotoksične in fenotipske lastnosti naravnih celic ubijalk, vendar učinek ni bil enoten. Najizrazitejši imunomodulatorni učinek smo opazili pri celični liniji PANC-1, kjer je pri določenih pogojih prišlo do povečanja izražanja katepsina C, perforina in grancima B, kar nakazuje povečan citotoksični potencial naravnih celic ubijalk. Pri celični liniji CAPAN-2 so bili učinki šibkejši, časovno bolj spremenljivi in pogosteje povezani s spremembami regulatorne molekule cistatina F. Analiza površinskih označevalcev ni pokazala enotnega premika proti izrazito aktiviranemu fenotipu, temveč kompleksno prilagoditev, odvisno od tumorske celične linije, časa izpostavitve in uporabljenega modela naravnih celic ubijalk. Pomembna ugotovitev raziskave je bila tudi razlika med celično linijo NK-92 in primarnimi naravnimi celicami ubijalkami. Celična linija NK-92 je pokazala bolj ponovljiv odziv, medtem ko so bile primarne celice izrazito odvisne od darovalca. Rezultati kažejo, da lahko gemcitabin posredno spremeni komunikacijo med tumorskimi in imunskimi celicami, vendar je ta učinek odvisen od lastnosti tumorskih celic, časovne dinamike odziva in biološke raznolikosti imunskih celic. Pancreatic cancer is one of the most aggressive malignancies and is characterized by a strongly immunosuppressive tumor microenvironment and poor response to treatment. Gemcitabine remains an important therapeutic option, but in addition to its direct cytotoxic effect on tumor cells, it may also influence their communication with immune cells. The aim of this master’s thesis was to investigate whether conditioned medium from pancreatic cancer cells previously treated with gemcitabine indirectly affects the phenotypic and functional characteristics of natural killer cells. In this study, the pancreatic cancer cell lines PANC-1 and CAPAN-2 were treated with gemcitabine, after which conditioned medium was prepared from their secreted factors. The conditioned medium was then applied to the NK-92 natural killer cell line and to primary natural killer cells isolated from two healthy donors. The expression of molecules involved in granule-mediated cytotoxicity was analyzed by western blotting, while the expression of surface markers related to activation, inhibition and adhesion was evaluated by flow cytometry. The results showed that conditioned medium from tumor cells affects natural killer cells, although the effect was not uniform. The most pronounced immunomodulatory effect was observed with the PANC-1 cell line, where under certain conditions increased expression of cathepsin C, perforin and granzyme B was detected, suggesting an enhanced cytotoxic potential of natural killer cells. In the CAPAN-2 cell line, the effects were weaker, more time-dependent and more often associated with changes in the regulatory molecule cystatin F. Analysis of surface markers did not reveal a uniform shift toward a clearly activated phenotype, but rather a complex adaptation depending on the tumor cell line, exposure time and the natural killer cell model used. An important finding of this study was also the difference between the NK-92 cell line and primary natural killer cells. The NK-92 cell line showed a more reproducible response, whereas primary cells displayed marked donor-dependent variability. Overall, the results indicate that gemcitabine can indirectly alter communication between tumor and immune cells, but this effect depends on tumor cell properties, response dynamics and the biological variability of immune cells. rak trebušne slinavke gemcitabin naravne celice ubijalke tumorsko mikrookolje citotoksičnost pancreatic cancer gemcitabine natural killer cells tumor microenvironment cytotoxicity true false false [V. Slatinek] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-09-04 07:45:09 2026-09-04 07:45:17 2026-09-08 11:26:04 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 616-097:577.27(043.2) 128036 290352643 33176.pdf 33176.pdf 1 3A92B56D2D2219FE9028C388CBDA328B f7f8ff336ac025a88dd1f5afeb283822280842c8879a20c45fe44ed863f51620 cf332528-a823-11f1-8bc5-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=245984 Fakulteta za farmacijo 0 0 0