magistrsko delo
Ajda Godec (Author), Maša Sinreih (Mentor), Miha Pavšič (Thesis defence commission member), Vera Župunski (Thesis defence commission member)

Abstract

Aldo-keto reduktaze tipa 1C (AKR1C) uvrščamo v proteinsko naddružino aldo-keto reduktaz, vključenih v metabolizem steroidnih hormonov, nevrosteroidov in prostaglandinov. Hkrati pa preko različnih mehanizmov sodelujejo pri razvoju kemorezistence, tako da so del razgradne poti kemoterapevtikov ali pa znižujejo obseg celičnega oksidativnega stresa, ki nastane kot posledica njihovega delovanja. Iz tega vidika encimi AKR1C predstavljajo zanimive molekularne tarče pri zdravljenju vrst raka odpornih na kemoterapevtike. V to skupino sodi tudi serozni karcinom jajčnikov visoke stopnje malignosti (HGSC), ki je najpogostejša vrsta raka jajčnikov in vodilni vzrok smrtnosti v skupini ginekoloških rakov. Namen magistrskega dela je bil proučiti potencialno inhibitorno delovanje 11 spojin na encime AKR1C1–C3 in raziskati vpliv izbranih spojin na viabilnost modelnih celičnih linij HGSC ter raziskati morebitne razlike v primerjavi s kontrolno celično linijo epitelija jajčnikov. V raziskavi smo uporabili 5 derivatov tetrahidronaftalen-1-ona (DTP-153, DTP-154, DTP-155, DTP-158, AD-13) in 6 derivatov 13α-estrona (AD-4, AD-5, DTP-036, DTP-026, DTP-150, DTP-150-KK). Inhibitorni učinek derivatov 13α-estrona in tetrahidronaftalen-1-ona smo preverjali z merjenjem encimske aktivnosti encimov AKR1C1–C3 v prisotnosti testne spojine pri končni koncentraciji 10 µM ali 100 µM, pri čemer smo spremljali reakcijo oksidacije 1-acenaftola ob sočasni redukciji koencima NAD+. Vpliv inhibitorjev na celično viabilnost smo določili s testom Alamar Blue, pri katerem smo kontrolni celični liniji jajčnikov (HIO-80) in celični liniji HGSC (COV362 in OVSAHO) za 48 ur izpostavili 11 sintetičnim inhibitorjem v različnih koncentracijah (10 µM in 50 µM). Derivati 13α-estrona so delovali kot boljši inhibitorji encimov AKR1C1–C3 kot tetrahidronaftalenski derivati. Pri 100 µM koncentraciji sta dve spojini iz skupine tetrahidronaftalenskih derivatov (DTP-154 in DTP-158) inhibirali encim AKR1C3 za več kot 50 %. Najbolje sta encim AKR1C2 inhibirali spojini DTP-036 in AD-4 z IC$_50$ vrednostmi 3,3 µM in 14,2 µM. Prav tako smo s programom AutoDock Vina umestili modele spojin DTP-036 in AD-4 v aktivno mesto encima AKR1C2. Nižjo eksperimentalno določeno vrednost IC50 za spojino DTP-036 lahko glede na napoved vezave povežemo s prisotnostjo dodatnih hidrofobnih interakcij. Ob dodatku testnih spojin DTP-153, DTP-155, AD-13 in DTP-150 pri koncentraciji 50 µM lahko opazimo statistično pomembno zmanjšanje deleža celične viabilnosti manj invazivne celične linije HGSC OVSAHO. Pri kontrolni celični liniji HIO-80 in celični liniji COV362 ni bilo opaženih statistično pomembnih vplivov na celično viabilnost. Na osnovi rezultatov lahko zaključimo, da smo identificirali dva nova selektivna inhibitorja encima AKR1C2 in štiri spojine, ki statistično pomembno zmanjšajo delež celične viabilnost manj invazivne celične linije HGSC OVSAHO

Keywords

aldo-keto reduktaze;AKR1C1-C3;encimska aktivnost;inhibicija;celična proliferacija;magistrska dela;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher: [A. Godec]
UDC: 577.15:618.11-006(043.2)
COBISS: 183355651 Link will open in a new window
Views: 52
Downloads: 11
Average score: 0 (0 votes)
Metadata: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Other data

Secondary language: English
Secondary title: 13α-estrone and tetrahydronaphthalen-1-one derivatives as AKR1C1–C3 inhibitors and their effect on high-grade serous ovarian cancer cell lines
Secondary abstract: Aldo-keto reductases type 1C (AKR1C), members of the aldo-keto reductase superfamily, are involved in the metabolism of steroid hormones, neurosteroids, and prostaglandins and contribute to chemoresistance, by either metabolizing chemotherapeutic agents or indirectly by eradicating the cellular stress caused by chemotherapeutics. These enzymes represent interesting drug targets for chemoresistant cancers. High-grade serous ovarian cancer (HGSOC) is the most common type of ovarian cancer and the leading cause of death out of all gynecological cancers. The aim of our master thesis was to investigate 11 compounds as inhibitors of aldo-keto reductase 1C (AKR1C) enzymes and their effects on proliferation of control and ovarian cancer cell lines (HGSOC). Six of the 11 compounds can be classified as estrane derivatives (AD-4, AD-5, DTP-036, DTP-026, DTP-150, DTP-150-KK), and the other 5 are tetrahydronaphtalen-1-one derivatives (DTP-153, DTP-154, DTP-155, DTP-158, AD-13). The inhibitory properties towards AKR1C1–C3 enzymes were measured at inhibitor concentrations of 10 and 100 µM with spectrophotometric detection of oxidation of the substrate 1-acenaphthenol in the presence of NAD + as coenzyme. The effect of inhibitors on cell viability was determined with Alamar Blue assay. The control ovarian cell line (HIO-80) and HGSOC cell lines (COV362 and OVSAHO) were exposed to 11 synthetic inhibitors at different concentrations (10 µM and 50 µM) for 48 hours. 13α-estrone derivatives were more effective inhibitors than tetrahydronaphtalen-1-one derivates. The two most active inhibitors of the AKR1C2 isoenzyme, DTP-036 and AD-4, had an IC50 value of 3.3 µM and 14.2 µM, respectively. At 100 µM concentration two of tetrahydronaphtalen-1-one derivates (DTP-154 in DTP-158) also showed more than 50% inhibition of AKR1C3. We also predicted the binding of AD-4 and DTP-036 into the binding pocket of AKR1C2 by using AutoDock Vina. The lower IC$_50$ value for DTP-036 correlates with the higher number of predicted hydrophobic interactions between DTP-036 and AKR1C2. Cell viability of the OVSAHO cells was significantly decreased when exposed to 50 µM DTP-153, DTP-155, AD -13, and DTP-150. There were no effects on control cell line HIO-80 and HGSOC cell line COV362. To conclude, we discovered new selective inhibitors of AKR1C2 isoform and four compounds that decrease viability of the HGSOC cell line OVSAHO
Secondary keywords: ovarian cancer;aldo-keto reductases;inhibition;cell viability;Encimi;Novotvorbe jajčnikov;Univerzitetna in visokošolska dela;
Type (COBISS): Master's thesis/paper
Study programme: 1000377
Thesis comment: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, smer Biokemija
Pages: 1 spletni vir (1 datoteka PDF (61 str.))
ID: 22766833