Utjecaj citarabina na stanični ciklus leukemijskih stanica U937

master's thesis

• Title (english): The effect of cytarabine on the cell cycle of U937 leukemia cells
• Author: Dora Matić
• Mentor: Dora Višnjić (mentor)
• Mentor: Inga Urlić (komentor)
• Committee member: Inga Urlić (predsjednik povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Committee member: Petra Korać (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2022-12-09, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Citarabin je jedan od standardno korištenih lijekova u liječenju akutne mijeloične leukemije.
Uočeno je da citarabin uzrokuje zastoj leukemijskih stanica u različitim fazama staničnog ciklusa.
Poznato je da zastoj u fazama S i G2/M aktivira kinazu Chk1 te da Chk1 inhibira CDK1, ali uloga
kinaze CDK1 u učincima citarabina nije opisana. Cilj ovoga rada je ispitati učinke citarabina na
stanični ciklus leukemijske linije U937 i kinazu CDK1, a hipoteza je da će citarabin zaustaviti
progresiju stanica U937 kroz stanični ciklus i inhibirati kinazu CDK1. Stanice tretirane citarabinom
obilježila sam propidij jodidom i odredila postotak stanica u pojedinoj fazi staničnog ciklusa
koristeći protočnu citometriju. Količinu kinaze CDK1 fosforilirane na tirozinu 15 (Tyr-15) u
stanicama koje su tretirane citarabinom tijekom 48 sati odredila sam metodom Western blot.
Rezultati su pokazali da citarabin smanjuje broj živih stanica u ovisnosti o koncentraciji te da u
višim koncentracijama uzrokuje zastoj u fazama S i G2/M. Zastoj staničnog ciklusa korelira s
fosforilacijom kinaze CDK1 na Tyr-15 što upućuje na ulogu CDK1 kao ciljne molekule signalnoga
puta ATR/Chk1 u učincima citarabina na leukemijske stanice U937.

Abstract (english)

Cytarabine is one of the standard drugs used in treating acute myeloid leukemia. It is noted that
cytarabine causes cell cycle arrest in leukemia cells in different phases of the cell cycle. Arrest in
the S and G2/M phases activates Chk1 which in turn inhibits CDK1, but the role of CDK1 in the
effects of cytarabine has not yet been described. The aim of this paper is to test the effect of
cytarabine on the cell cycle of U937 cells and kinase CDK1, while the hypothesis is that cytarabine
will halt progression of U937 cells through the cell cycle and inhibit CDK1. I marked the cells
treated with cytarabine with propidium iodine and determined the percentage of cells in each phase
of the cell cycle using flow cytometry. To determine the level of CDK1 phoshorylated on Tyr-15
in cells treated with cytarabine for 48 hours I used Western blot analysis. The results show that
cytarabine lowers the number of living cells in a dose dependent manner and in higher
concentrations causes cell cycle arrest in the S and G2/M phase. Cell cycle arrest correlates with
the phoshorylation of CDK1 on Tyr-15 which indicates the role of CDK1 as a key molecule in the
ATR-Chk1 signaling pathway in the effects of cytarabine on U937 leukemia cells.

• Keywords: CDK1; Chk1; AML
• Keywords (english): CDK1; Chk1; AML
• Language: croatian
• URN:NBN: urn:nbn:hr:217:521857
• 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
• Created on: 2022-12-20 13:00:13