1. Metode določevanja cirkulirajočih tumorskih celic raka dojke na Onkološkem inštitutu LjubljanaTanja Jesenko, Simona Miceska, Živa Pišljar, Maja Čemažar, Veronika Kloboves-Prevodnik, Cvetka Grašič-Kuhar, 2024, published scientific conference contribution Keywords: rak dojke, patologija, onkologija
Published in DiRROS: 25.02.2026; Views: 127; Downloads: 24
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2. Protitumorsko delovanje utišanja transkripcijskega faktorja E2F1 na modelu humanega kolorektalnega karcinoma : in vitro in in vivo spoznanjaSimona Kranjc Brezar, Tanja Jesenko, Monica Cazzato, Živa Pišljar, Boštjan Markelc, Tim Božič, Gabriele Grassi, Maja Čemažar, 2024, published scientific conference contribution abstract (invited lecture) Keywords: rak debelega črevesa in danke, eksperimentalna onkologija, onkologija
Published in DiRROS: 25.02.2026; Views: 116; Downloads: 19
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3. Vzpostavitev in karakterizacija mišjega tumorskega modela za raziskave s HPV povezanega raka glave in vratuŽiva Pišljar, Maja Čemažar, Boštjan Markelc, Gregor Serša, Simona Kranjc Brezar, Tanja Jesenko, 2024, published scientific conference contribution abstract Keywords: elektroprenos genov, eksperimentalna onkologija, onkologija
Published in DiRROS: 25.02.2026; Views: 124; Downloads: 19
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4. Chemokine CCL5 overexpression combined with radiotherapy modulates Th1-mediated immune response and leads to significant tumor growth delay in mouse tumor modelsTim Božič, Iva Šantek, Živa Pišljar, Simona Kranjc Brezar, Gregor Serša, Boštjan Markelc, Maja Čemažar, 2026, original scientific article Abstract: This study investigated the antitumor efficacy of chemokine CCL5 gene therapy using gene electrotransfer (GET) in combination with radiotherapy (RT) in solid murine tumors CT26 and 4T1. In vitro, CT26 and 4T1 tumor cells transfected with plasmid DNA (pDNA) encoding CCL5 induced migration of RAW264.7 macrophages. In vivo, CCL5 overexpression achieved via GET of pDNA encoding CCL5 led to increased splenocyte infiltration in dorsal window chamber models. When combined with RT, GET of pDNA encoding CCL5 shifted the tumor cytokine profile toward a proinflammatory state, with elevated Ifn-γ, Cxcl9, Cxcl10, and Il-12α. Although CD8 + and CD4 + T cells were reduced post-treatment, due to radiation-induced cell death, the combination of GET of pDNA encoding CCL5 and RT significantly delayed tumor growth in both models. In 4T1 tumors, this delay was also significant compared to the equivalent treatment with GET of control pDNA. These findings support GET of pDNA encoding CCL5 combined with RT as a strategy to enhance immune-mediated tumor control.
Keywords: chemokines, gene electrotransfer, gene therapy, mouse, radiotherapy
Published in DiRROS: 13.02.2026; Views: 339; Downloads: 47
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5. Complementary detection strategies for circulating tumor cells in breast cancer: clinical implications of combining immunofluorescence and cytopathological stainingTanja Jesenko, Cvetka Grašič-Kuhar, Živa Pišljar, Simona Miceska, Veronika Kloboves-Prevodnik, Maja Čemažar, 2025, original scientific article Abstract: Background: Circulating Tumor Cells (CTCs) serve as important biomarkers for disease monitoring and treatment response in patients with metastatic breast cancer. Their detection remains challenging because of their low abundance, phenotypic diversity and non-standardized mode of detection. Cytopathological Giemsa and Immunofluorescence (IF) staining can offer complementary approaches for CTC characterization. Giemsa staining enables assessment of cellular morphology, while IF allows for marker-specific identification, together providing a more comprehensive and accurate evaluation of CTCs. Methods: We developed an IF staining protocol with antibodies against Cytokeratin (CK), vimentin (VIM), and Cluster of Differentiation 45 (CD45) to distinguish epithelial, mesenchymal, hybrid and hematopoietic cells for CTC detection and characterization and compared it with cytopathologic method of detection via Giemsa staining with regard to CTC detection rates and morphological detail. Results: Study was performed on the samples of 29 heavily pretreated patients with metastatic breast cancer (median duration of metastatic disease 19.4 months). Giemsa staining enabled the detection of a higher number of CTCs compared to our IF protocol. Lower detection rate was potentially due to the loss of fragile or loosely adherent cells during methanol fixation and IF staining. Additionally, in IF-stained samples, some CTCs presented faint nuclear signals, potentially impairing their recognition. The IF staining supported the identity of CTCs detected on Giemsa-stained slides by employing a three-color antibody panel-based approach and allowed detailed phenotypic discrimination and structural analysis of CTCs, including the identification of a distinctive CK polarization pattern suggestive of a transitional state during intravasation. Conclusion: Giemsa and IF may thus be complementary rather than mutually exclusive and relying on a single detection approach could underestimate the true CTC burden. An integrative strategy combining both techniques may offer a more comprehensive view of CTC populations in metastatic breast cancer, thereby enhancing diagnostic precision.
Keywords: biomarkers, breast cancer, circulating tumor cells, cytopathological detection
Published in DiRROS: 05.12.2025; Views: 928; Downloads: 101
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6. Razvoj raziskav cirkulirajočih tumorskih celic pri raku dojk na Onkološkem inštitutu LjubljanaTanja Jesenko, Cvetka Grašič-Kuhar, Živa Pišljar, Simona Miceska, Veronika Kloboves-Prevodnik, Maja Čemažar, 2024, review article Abstract: Cirkulirajoče tumorske celice (CTC) so postale pomemben biološki označevalec pri raku dojk, saj omogočajo vpogled v razvoj in napredovanje razsejane bolezni ter spremljanje odziva na zdravljenje. Zaradi njihove izjemne redkosti in kompleksnosti sestave krvi, v kateri se nahajajo, sta njihova izolacija in karakterizacija velik izziv. Posebne metode izolacije omogočajo obogatitev CTC iz vzorca krvi in olajšajo nadaljnjo analizo. Na Onkološkem inštitutu Ljubljana smo leta 2018 začeli s prvimi koraki v smeri razvoja preproste metode za izolacijo in karakterizacijo CTC, ki bi omogočala prepoznavanje teh celic s citopatološkimi analizami. Ocenili smo dve različni metodi izolacije CTC pri bolnicah z rakom dojk, ki temeljita na različnih pristopih. Prva metoda temelji na bioloških lastnostih celic, kot je izražanje epitelijskega označevalca celične adhezije (EpCAM), medtem ko druga metoda temelji na fizikalnih lastnostih CTC, kot sta večja velikost in stisljivost v primerjavi z drugimi krvnimi celicami. Ugotovili smo, da je fizikalna metoda primernejša, saj omogoča izolacijo večjega števila morfološko ohranjenih CTC in tudi skupkov CTC. Po izolaciji pripravimo citološke preparate, ki jih nato opredelimo s citopatološko analizo in dodatnimi imunocitokemičnimi ter imunofluorescenčnimi barvanji. Na ta način lahko trenutno določimo število CTC in skupkov CTC v krvi, ocenimo njihovo morfološko ohranjenost ter prepoznamo njihov fenotip. Poleg preučevanja vzorcev posamičnih CTC in skupkov CTC v okviru trenutno potekajočih kliničnih raziskav in načrtovane vzpostavitve translacijske platforme na mišjih modelih, pa v prihodnosti želimo nabor raziskav CTC še razširiti na genomsko in transkriptomsko analizo.
Keywords: cirkulirajoče tumorske celice, rak dojk, eksperimentalna onkologija
Published in DiRROS: 21.05.2025; Views: 633; Downloads: 208
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7. Morphological features of breast cancer circulating tumor cells in blood after physical and biological type of isolationTanja Jesenko, Živa Pišljar, Cvetka Grašič-Kuhar, Maja Čemažar, Urška Matkovič, Simona Miceska, Jerneja Varl, Anamarija Kuhar, Veronika Kloboves-Prevodnik, 2021, original scientific article Abstract: Background. Circulating tumor cells (CTCs) have become an important biomarker in breast cancer. Different iso-lation tech-niques based on their biological or physical features were established. Currently, the most widely used methods for visualization after their separation are based on immunofluorescent staining, which does not provide the information on the morphology.Materials and methods. The aim of this study was to evaluate how two different separation techniques affect cell morphology and to analyse cell morphology with techniques used in routine cytopathological laboratory. A direct side-by-side comparison of physical (Parsortix%) and biological (MACS%) separation technique was performed.Results. In the preclinical setting, both isolation techniques retained the viability and antigenic characteristics of MCF7 breast cancer cells. Some signs of degeneration such as cell swelling, cytoplasmic blebs, villous projections and vacuolization were observed. In metastatic breast cancer patient cohort, morphological features of isolated CTCs were dependent on the separation technique. After physical separation, CTCs with preserved cell morphology were detected. After biological separation the majority of the isolated CTCs were so degenerated that their identity was difficult to confirm.Conclusions. Taken together, physical separation is a suitable technique for detection of CTCs with preserved cell morphology for the use in a routine cytopathological laboratory.
Keywords: circulating tumor cells, breast cancer, morphology
Published in DiRROS: 22.07.2024; Views: 1284; Downloads: 812
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8. Razvoj in karakterizacija mišjega modela za študije HPV-pozitivnega raka glave in vratuŽiva Pišljar, Maja Čemažar, Boštjan Markelc, Andrej Cör, Gregor Serša, Simona Kranjc Brezar, Tanja Jesenko, 2023, published scientific conference contribution abstract Keywords: miši, rak glave in vratu, gensko zdravljenje
Published in DiRROS: 19.06.2023; Views: 1388; Downloads: 687
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9. Nižje število cirkulirajočih tumorskih celic v poznih linijah razsejanega raka dojkSilvester Jernej, Klara Geršak, Marina Mencinger, Tanja Ovčariček, Tanja Jesenko, Živa Pišljar, Simona Miceska, Maja Čemažar, Veronika Kloboves-Prevodnik, Cvetka Grašič-Kuhar, 2022, published professional conference contribution abstract Keywords: onkologija, rak dojke, kemoterapija
Published in DiRROS: 03.02.2023; Views: 2300; Downloads: 498
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10. Morfološke značilnosti cirkulirajočih tumorskih celic raka dojk v krvi po izolaciji na podlagi njihovih fizikalnih ali bioloških lastnostiSimona Miceska, Tanja Jesenko, Živa Pišljar, Cvetka Grašič-Kuhar, Maja Čemažar, Urška Matkovič, Jerneja Varl, Anamarija Kuhar, Veronika Kloboves-Prevodnik, 2022, published scientific conference contribution abstract Keywords: citologija, diagnostika, onkologija
Published in DiRROS: 27.01.2023; Views: 1891; Downloads: 449
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