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Isolation, cultivation and characterization of cells originating from tumor and tumor margin of patients with basal cell carcinoma in vitro

dc.contributor.advisorPetrović, Milan B.
dc.contributor.otherMilašin, Jelena
dc.contributor.otherDanilović, Vesna
dc.contributor.otherJovanović-Radivojević, Svetlana
dc.contributor.otherJezdić, Zoran
dc.creatorMilošević, Maja
dc.date.accessioned2020-02-27T11:21:44Z
dc.date.available2020-02-27T11:21:44Z
dc.date.available2020-07-03T09:15:32Z
dc.date.issued2019-10-22
dc.identifier.urihttp://eteze.bg.ac.rs/application/showtheses?thesesId=7296
dc.identifier.urihttps://nardus.mpn.gov.rs/handle/123456789/12099
dc.identifier.urihttps://fedorabg.bg.ac.rs/fedora/get/o:21075/bdef:Content/download
dc.identifier.urihttp://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=51882767
dc.description.abstractHomeostaza kože direktno zavisi od mehanizma kontinuiranog obnavljanja ćelijske populacije kože i njene sposobnosti regeneracije. U epidermisu je prisutna posebna populacija ćelija zadužena za proces samoobnavljanja kože, tzv. epidermalne matične ćelije koje imaju sposobnost proliferacije, migracije, diferencijacije i programirane ćelijske smrti. Ukoliko dođe do narušavanja ravnoteže između ovih procesa, može doći do maligne transformacije zdrave matične ćelije u kancersku matičnu ćeliju. Poznato je da za hemorezistenciju, metastaziranje i recidiviranje tumora odgovornost nosi posebna populaciju tumorskih ćelija koje se nazivaju kancerske matične ćelije. Istraživanja pokazuju da bazocelularni karcinom kože vodi poreklo od matičnih ćelija ispupčenja folikula dlake, infundibuluma dlake i interfolikularnog epidermisa. Bazocelularni karcinom (BCK) je najčešći maligni tumor kože, pa i ako retko metastazira karakteriše ga invazivni rast pri čemu infiltriše okolne strukture i dovodi do ozbiljnih funkcionalnih i estetskih defekata. U literaturi se kao jedan od ključnih faktora odgovornih za recidiviranje navodi neadekvatna širina margine resekcije tumora. Istraživanja su pokazala da je za primarne BCK na koži lica veličine do 20 mm preporučena širina margine 3 mm. Cilj ovog istraživanja bio je identifikacija i karakterizacija subpopulacije ćelija sa osobinama matičnosti izolovanih iz BCK-a i bliske margine od 3 mm, ispitivanjem proliferativnog, klonogenog i migratornog potencijala, analizom ekspresije embrionalnih, mezenhimskih i tumorskih markera, analizom biohemijske strukture ćelija, ispitivanjem sposobnosti diferencijacije u više ćelijskih linija kao i stepena otpornosti ćelija na dejstvo citostatika. U istraživanju je učestvovalo 13 pacijenata sa primarnim bazocelularnim karcinomom na koži lica veličine do 20 mm. Nakon detaljno uzete anamneze i potpisanog informisanog pristanka pristupalo se hirurškoj eksciziji tumorske promene, koja se smatra zlatnim standardom u terapiji BCK-a. Iz svih ekscidiranih uzorka za izolaciju ćelija uzet je deo tumorskog tkiva, tkiva bliske margine (udaljene 3 mm od makroskopske granice tumora) i deo zdrave kože obuhvaćen oblikom resekcije (udaljen od makroskopske granice tumora više od 5 mm), dok je preostali deo tkiva poslat na histopatološku analizu. Za karakterizaciju ćelija u kulturi korišćeni su testovi proliferacije, migracije, formiranja kolonija, sfera i ramanska spektroskopija. Markeri kancerskih matičnih ćelija (KMĆ) analizirani su ,,Real-Time PCR” metodom i protočnom citometrijom...sr
dc.description.abstractrenewal and regeneration and the epidermis harbors a special cell population responsible for the process of self-renewal and regeneration of the skin, so called epidermal stem cells that have the ability to proliferate, migrate, differentiate, and undergo programmed cell death. If imbalance between these processes occurs, malignant transformation of healthy stem cell into the cancerous stem cell may happen. It is also known that a specific population of tumor cells called cancer stem cells is responsible for the hemoresistance, metastasis, and recurrence. Studies have shown that skin basal cell carcinoma is caused by stem cells of hair follicle, hair infundibulum and interfollicular epidermis. Basocellular carcinoma (BCC) is the most common malignant tumor of the skin, but it rarely gives metastases It is characterized by invasive growth, infiltrating surrounding structures and leading to serious functional and aesthetic defects. In the literature, inadequate margins are indicated as one of the key factors responsible for tumor recurrences. Studies have shown that for the primary facial skin BCC of less than 20 mm a margin of 3 mm is recommended. The aim of this study was to identify and characterize the cell subpopulation with stem characteristics whithin BCC and its margins (3 mm close) by testing proliferative, clonogenic and migratory potential, by analyzing the expression of embryonic, mesenchymal and tumor markers, by analizing biochemical structure of cells, by examining the differentiation potential of generated cell lines and by testing the degree of chemo-resistance. The study included 13 patients with primary BCC of face skin of less than 20 mm. Following a detailed anamnesis and a signed informed consent, the surgical excision which is considered a gold standard in BCC therapy, was done. A portion of the tumor tissue, of close margin (3 mm from the macroscopic tumor limit), and of healthy skin included in the resecion (at least 5mm from the macroscopic tumor limit), were obtained from all the patients. The remaining of the tissue was subjected to histopathological analysis. For cell phenotype characterization, tests of proliferation, colony formation, scratch assay, and sphere formation were used...en
dc.formatapplication/pdf
dc.languagesr
dc.publisherУниверзитет у Београду, Стоматолошки факултетsr
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/175075/RS//
dc.rightsopenAccessen
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/
dc.sourceУниверзитет у Београдуsr
dc.subjectbazocelularni karcinomsr
dc.subjectbasal cell carcinomaen
dc.subjectkancerske matične ćelijesr
dc.subjecthirurška marginasr
dc.subjectdiferencijacijasr
dc.subjectgenska ekspresijasr
dc.subjectklonogenostsr
dc.subjecthemorezistentnostsr
dc.subjectcancer stem cellsen
dc.subjectneoplastic cellsen
dc.subjectsurgical marginen
dc.subjectdifferentiationen
dc.titleIzolacija, kultivizacija i karakterizacija ćelija poreklom iz tumora i tumorske margine pacijenata sa bazocelularnim karcinomom u in-vitro uslovimasr
dc.title.alternativeIsolation, cultivation and characterization of cells originating from tumor and tumor margin of patients with basal cell carcinoma in vitroen
dc.typedoctoralThesisen
dc.rights.licenseBY-NC
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/16445/IzvestajKomisije22212.pdf
dc.identifier.fulltexthttps://nardus.mpn.gov.rs/bitstream/id/16444/Disertacija.pdf
dc.identifier.fulltexthttps://nardus.mpn.gov.rs/bitstream/id/16445/IzvestajKomisije22212.pdf
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/16444/Disertacija.pdf
dc.identifier.rcubhttps://hdl.handle.net/21.15107/rcub_nardus_12099


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