Proučevanje sinteze androgenov v modelnih celičnih linijah raka endometrija

Šturm, Lea (author), Gobec, Martina (mentor), Sinreih, Maša (co-mentor)

Rak endometrija je najpogostejše ginekološko obolenje, ki se večinoma pojavi po menopavzi. Glede na histološke značilnosti ga delimo na tip 1 in tip 2. Tip 1 je pogostejši tip raka, nizkega gradusa, pozitiven na hormonske receptorje ter predvsem povezan z debelostjo in metabolnim sindromom. Tip 2 je neendometrioiden, pogosto negativen na hormonske receptorje in ima večjo verjetnost metastaziranja in slabšo prognozo. Androgeni v rodni dobi nastajajo v nadledvični žlezi ter jajčnikih. Nadledvična žleza sintetizira malo aktivnih androgenov, večinoma sintetizira prekurzorje, ki se v perifernih tkivih pretvorijo v aktivne androgene. Ti nato z vezavo na receptor za androgene izzovejo fiziološki odgovor. Koncentracija klasičnih androgenov s starostjo v krvnem obtoku žensk pada za razliko od 11-oksi androgenov, katerih koncentracija se s starostjo ne zmanjšuje. Vloga estrogenov je v patologiji raka endometrija dobro proučena v nasprotju z vlogo androgenov. Estrogeni stimulirajo proliferacijo epiteljiskih celic v endometriju, kar vodi v večjo možnost kopičenja genetskih napak. V primeru, da delovanju estrogenov ne nasprotuje progesteron, se verjetnost za nastanek raka endometrija poveča. Vpliv androgenov na nastanek in napredovanje raka endometrija ni še povsem znan. Epidemiološke raziskave povezujejo povečano koncentracijo androgenov v krvnem obtoku žensk z večjo verjetnostjo nastanka raka endometrija, predvsem na račun aromatizacije androgenov v maščobnem tkivu. Raziskave na celičnih linijah kažejo na to, da imajo androgeni z vezavo na receptor za androgene zaviralen učinek na rast celic in bi tako lahko imeli pri razvoju in napredovanju raka endometrija zaščitno vlogo. Namen magistrskega dela je proučiti metabolizem androgenov v modelnih celičnih linijah raka endometrija ter kontrolni celični liniji. Celične linije smo tretirali z različnimi androgeni, ki predstavljajo prekurzorje v sintezi aktivnih klasičnih in 11-oksi androgenov. Metabolite smo ekstrahirali z ekstrakcijo trdno-tekoče ali ekstrakcijo tekoče-tekoče in jih nato določili s tekočinsko kromatografijo, sklopljeno s tandemsko masno spektrometrijo (LC-MS/MS). Na podlagi rezutatov smo ugotovili razlike v metabolizmu med posameznimi celičnimi linijami. Za najbolj metabolno aktivni celični liniji sta se izkazali RL95-2 in Ishikawa, pri katerih smo zaznali največ aktivnih androgenov. Ugotovili smo, da se aktivni androgeni v modelnih celičnih linijah raka endometrija lahko tvorijo iz prekurzorjev, kot so dihidroepiandrosteron (DHEA), dihidroepiandrosteron sulfat (DHEAS), androstendion (A4), 11-hidroksi androstendion (11OHA4) in 11-ketoandrostendion (11KA4).

• Language: Slovenian
• Keywords: androgeni, metabolizem, rak endometrija
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2023
• PID: 20.500.12556/RUL-145676
• Publication date in RUL: 07.05.2023

Secondary language

• Language: English
• Title: A study of androgen synthesis in endometrial cancer model cell lines

Abstract

Endometrial carcinoma (EC) is one of the most common gynecological malignancy, which mostly occurs after menopause. Depending on the histological characteristics, EC has been divided into type 1 and type 2. Type 1 is usually low grade, hormone-receptor positive and associated with obesity and metabolic syndrome. Type 2 is associated with non-endometrioid histology, worse prognosis and it's usually hormone-receptor negative and more likely to metastasize. Androgens are produced by the adrenal gland and gonads. The adrenal gland synthesizes low amounts of active androgens, mostly precursors that are converted into active androgens in peripheral tissues. Levels of classical androgens decline after menopause, but 11-oxyandrogens levels remains unchanged during the menopausal years. It is well known that estrogens play pivotal role in the developement and progression of endometrial carcinoma. Estrogens promote cell proliferation, which leads to a greater chance of DNA replication errors. Excesssive exposure to estrogens and low progesterone levels are considered to increase the risk of EC. The impact of androgens on the formation and progression of endometrial cancer is not yet fully understood. Epidemiological evidences suggegest that there is increased risk of EC associated with elevated concentrations of androgens in women bloodstream. Studies on cell lines have shown that AR-dependent signalling inhibits cell proliferation and that androgens could play a protective role in the development and progression of EC. The aim of the master thesis is to study the metabolism of androgens in endometrial cancer model cell lines and a control cell line. Cell lines were treated with different androgens, which represent precursors in the synthesis of active classical and 11-oxyandrogens. Metabolites were extracted by solid-liquid or liquid-liquid extraction and then detected by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Based on the results, we found differences in metabolism between model cell lines. The most metabolically active cell lines were RL95-2 and Ishikawa. From our research we concluded that in the endometrial cancer model cell lines active androgens can be formed from the precursors such as dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), androstendione (A4), 11-hydroxyandrostendione (11OHA4) and 11-ketoandrostendione (11KA4).

• Keywords: androgens, endometrial cancer, metabolism