Utjecaj derivata primakina na modulaciju aktivnosti transportera ABCG2

master's thesis

• Title (english): The influence of primaquine derivatives on the modulation of transporter ABCG2 activity
• Author: Karla Carević
• Mentor: Marijeta Kralj (mentor)
• Mentor: Maja Matulić (sumentor)
• Committee member: Maja Matulić (predsjednik povjerenstva)
• Committee member: Mirna Ćurković Perica (član povjerenstva)
• Committee member: Dubravka Hranilović (član povjerenstva)
• Committee member: Antun Alegro (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2020-01-24, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Otpornost tumorskih stanica na kemoterapiju velik je problem u liječenju tumora. Najviše istražen mehanizam otpornosti jest povećano izbacivanje lijeka iz stanica pomoću transportera ABC. Oni normalno sudjeluju u izbacivanju različitih toksina i ksenobiotika te na taj način štite stanicu od štetnih utjecaja. Međutim, njihova prekomjerna ekspresija na površini tumorskih stanica vodi do otpornosti te smanjenog odgovora na liječenje. ABCG2 je protein veličine 72 kDa iz obitelji transportera ABC koji ima širok raspon supstrata, među kojima su i mnogi lijekovi. S obzirom da ABCG2 može značajno modulirati učinkovitost liječenja, važno je istražiti njegove interakcije sa spojevma. U ovome radu ispitivali smo djelovanje šest derivata primakina iz skupine fumardiamida primakina i halogenanilina i bis-urea derivata primakina, sintetiziranih kao potencijalni antimalarici i/ili protutumorski lijekovi na Farmaceutsko-biokemijskom fakultetu u Zagrebu, na aktivnost transportera ABCG2. Kao model koristili smo staničnu liniju akutne mijeloidne leukemije ljudi bez (PLB) i s povećanom ekspresijom ABCG2 (PLB-G2), koja pokazuje otpornost prema različitim supstratima ove pumpe, poput mitoksantrona. Ispitivali smo antiproliferativna svojstva spojeva testom MTT, dok smo inhibiciju ABCG2 ispitali metodom nakupljanja supstrata Hoechst 33342 i testom vezanja konformacijski osjetljivog protutijela 5D3, protočnom citometrijom, uz korištenje Ko143 kao pozitivne kontrole. Spojeve s najjačom aktivnosti testirali smo kombiniranim tretmanom s mitoksantronom, prateći eventualno pojačanje djelovanja mitoksantrona u otpornim stanicama.

Abstract (english)

The resistance of tumor cells to chemotherapy is a major problem in the treatment of tumors. The most investigated mechanism of resistance is the increased excretion of the drug from cells by the ABC transporter. They normally participate in the excretion of various toxins and xenobiotics, thus protecting the cell from adverse effects. However, their overexpression on the surface of the tumor cells leads to resistance and a reduced response to treatment. ABCG2 is a 72 kDa protein from the ABC transporter family that has a wide variety of substrates, including many known drugs. Given that ABCG2 can significantly modulate treatment efficacy, it is important to investigate its interactions with compounds. In this study, we investigated the effects of six primaquine derivatives (primaquine and halogenaniline fumardiamides and bis-urea primaquine derivatives), synthesized as potential antimalarials and/or anticancer drugs at the Faculty of Pharmacy and Biochemistry in Zagreb, on ABCG2 transporter activity. As a model, we used a cell line of acute myeloid leukemia without (PLB) and with increased expression of ABCG2 (PLB-G2), which shows resistance to various substrates of this pump, such as mitoxantrone. We tested the antiproliferative properties of the compounds using the MTT assay, while the inhibition of ABCG2 was tested with the Hoechst 33342 substrate accumulation assay and with 5D3 shift assay, by flow cytometry, using Ko143 as a positive control. The compounds with the strongest activity were tested with a combination treatment with mitoxantrone, to verify possible enhancement of mitoxantrone activity in resistant cells.

• Keywords: derivati primakina; transporter ABCG2; inhibicija; rak
• Keywords (english): primaquine derivatives; ABCG2 transporter; inhibition; cancer
• Language: croatian
• URN:NBN: urn:nbn:hr:217:896695
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2022-01-24
• Created on: 2020-06-16 09:30:39