magistrsko delo
Nika Požun (Author), Nada Žnidaršič (Reviewer), Mateja Erdani-Kreft (Mentor)

Abstract

Rak sečnega mehurja je deseto najpogostejše rakavo obolenje na svetu. Uporaba humane amnijske membrane (hAM) predstavlja obetaven pristop za zdravljenje raka. Z uporabo metodoloških pristopov qPCR, prenos western in imunooznačevanje, smo v normalnih in rakavih urotelijskih celicah proučili učinek homogenata hAM na izražanje izbranih genov ter prisotnost in znotrajcelično razporeditev proteinov, ki so povezani z avtofagijo. Rezultati nakazujejo, da je homogenat hAM v normalnih in rakavih urotelijskih celicah nekoliko zmanjšal izražanje izbranih genov (GOLPH3, BECN1, MAP1LC3B in SIRT1). Pri čemer smo statistično razliko dokazali samo pri izražanju GOLPH3 v rakavih urotelijskih celicah. Rezultati kažejo, da je homogenat hAM v rakavih celicah rahlo zmanjšal izražanje proteinov GOLPH3 in Beclin1 ter zvišal izražanje proteina LC3B II, vendar ne statistično značilno. Imunooznačevanje je rezultate preslikave western za GOLPH3 in Beclin1 potrdilo, ne pa tudi za LC3B, kjer je imunooznačevanje pokazalo nižje izražanje LC3B v tretiranih rakavih celicah. Ugotovili smo tudi, da homogenat hAM v rakavih urotelijskih celicah vpliva bodisi na povečanje razporeditve GOLPH3 v Golgijevem aparatu, bodisi vpliva na GOLPH3, da ostane prisoten v Golgijevem aparatu. Medtem ko v normalnih urotelijskih celicah sproži prerazporeditev GOLPH3 po večjem delu citoplazme. Na osnovi rezultatov sklepamo, da homogenat hAM preko različnih mehanizmov lahko inhibira avtofagijo v rakavih urotelijskih celicah, ne pa tudi v normalnih urotelijskih celicah. Menimo, da vpliv homogenata hAM na avtofagijo predstavlja potencialno področje nadaljnih raziskav v povezavi z zdravljenjem raka sečnega mehurja.

Keywords

humana amnijska membrana;homogenat;normalne in rakave urotelijske celice;avtofagija;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [N. Požun]
UDC: 577.2:616.62-006(043.2)
COBISS: 169212675 Link will open in a new window
Views: 35
Downloads: 10
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Other data

Secondary language: English
Secondary title: Effect of human amniotic membrane homogenate and defactinib on autophagy of urothelial cancer cells
Secondary abstract: Bladder cancer is the tenth most common cancer in the world. hAM homogenate is a promising approach to treat it. We used different methods (qPCR, Western blotting and immunolabelling) to investigate how hAM homogenate affects the expression of selected genes, as well as the expression and presence of selected proteins related to autophagy, in normal and cancer urothelial cells. The results indicate that hAM homogenate slightly decreased the expression of some genes (GOLPH3, BECN1, MAP1LC3B and SIRT1) in normal and cancer urothelial cells. We showed a statistical difference only in the expression of GOLPH3 in cancer urothelial cells. Furthermore, the results show that hAM homogenate slightly decreased the expression of proteins GOLPH3 and Beclin1 and increased the expression of LC3B II protein in cancer cells, but not statistically significant. Immunofluorescence confirmed the results of Western blotting results for GOLPH3 and Beclin1, but not for LC3B, with immunofluorescence showing lower expression of LC3B in treated cancer cells. We also found that hAM homogenate either increases the redistribution of GOLPH3 to the Golgi apparatus in cancer urothelial cells or influences GOLPH3 to remain present in the Golgi apparatus. Whereas in normal urothelial cells it triggers redistribution to the larger part of the cytoplasm. Based on the results, we conclude that hAM homogenate can inhibits autophagy in cancer urothelial cells by different mechanisms, but not in normal urothelial cells. We believe that the effect of hAM homogenate on autophagy represents a potential area for further research related to the treatment of bladder cancer.
Secondary keywords: human amniotic membrane;homogenate;normal and cancer urothelial cells;autophagy;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Oddelek za biologijo, Magistrsko delo magistrskega študijskega programa 2. stopnje Molekulske in funkcionalne biologije
Pages: 1 spletni vir (1 datoteka PDF (XIII, 72 str.)
ID: 20278484