Kombinirani učinak inhibitora tirozin kinaza i protutumorskih lijekova u stanicama melanoma čovjeka

master's thesis

• Title (english): Combination effect of tyrosine kinase inhibitors and antitumor drugs in human melanoma cells
• Author: Nebojša Vujnović
• Mentor: Andreja Ambriović Ristov (mentor)
• Mentor: Inga Marijanović (mentor)
• Committee member: Inga Marijanović (predsjednik povjerenstva)
• Committee member: Sofia Ana Blažević (član povjerenstva)
• Committee member: Tomislav Ivanković (član povjerenstva)
• Committee member: Petra Korać (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2017-02-27, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Stanice melanoma čovjeka izrazito su otporne na djelovanje protutumorskih lijekova.
Stoga je važno razviti nove terapije, a jedna od njih je ciljana terapija inhibitorima tirozin kinaza
i mogućnost njihove kombinacija s protutumorskim lijekovima. Stoga je u ovom radu na
modelu četiri stanične linije melanoma čovjeka istražena mogućnost liječenja melanoma
kombinacijom dasatiniba s protutumorskim lijekovima paklitakselom, vinkristinom ili
cisplatinom. Preživljenje stanica nakon izlaganja navedenim spojevima ili njihovim
kombinacijama određeno je testom MTT, dok je računanjem kombinacijskog indeksa
procijenjen zajednički učinak. Pokazano je da su stanice RPMI-7951 i MeWo osjetljive na
paklitaksel, vinkristin i cisplatinu, za razliku od stanica SK-MEL-3 koje pokazuju otpornost na
sva tri protutumorska lijeka i stanica A375 koje su otporne na paklitaksel i vinkristin, a osjetljive
na cisplatinu. Stanice RPMI-7951 pokazale su se osjetljivim na dasatinib, a njegova
kombinacija sa paklitakselom, vinkristinom i cisplatinom daje antagonistički učinak. Kod
stanica A375 uočena je slaba osjetljivost na dasatinib, a kombinacija sa cisplatinom pokazala
je sinergistički učinak. Na temelju određivanja količine Src/pSrc(Y418) metodama SDS-PAGE
i Western blot i rezultata preživljenja nakon utišavanja kinaze Src u stanicama RPMI-7951
zaključeno je da pSrc(Y418) nije meta dasatiniba te da su potrebna daljnja istraživanja u cilju
otkrivanja biomarkera za predviđanje osjetljivost stanica melanoma na dasatinib.

Abstract (english)

Human melanoma cells are highly resistant to antitumor drugs. Therefore, it is important
to develop new therapies such as targeted therapy with tyrosine kinases inhibitors, as well as
combining those inhibitors with antitumor drugs. In this research the potential of treating
melanoma with the combination of dasatinib and antitumor drugs paclitaxel, vincristine or
cisplatin in four human melanoma cell lines was investigated. Cell survival was determined by
MTT test, and combined effect was assessed by calculation of combination index. Cell lines
RPMI-7951 and MeWo were shown to be sensitive to paclitaxel, vincristine and cisplatin, SKMEL-
3 cells were shown to be resistant, while A375 cells were shown to be resistant to
paclitaxel and vincristine but sensitive to cisplatin. RPMI-7951 cells demonstrated sensitivity
to dasatinib, and its combination with paclitaxel, vincristine and cisplatin was shown to have
antagonistic effect. A375 cells demonstrated a mild sensitivity to dasatinib, and its combination
with cisplatin was shown to have a synergistic effect. On the basis of the amount of
Src/pSrc(Y418) determined by SDS-PAGE and Western blot and the survival of RPMI-7951
cells upon silencing Src kinase, it is concluded that pSrc(Y418) is not a dasatinib target,
emphasizing the need for identification of biomarkers that might predict sensitivity to dasatinib
in melanoma cells.

• Keywords: melanom; pSrc(Y418); dasatinib; cisplatina; paklitaksel; vinkristin
• Keywords (english): melanoma; pSrc(Y418); dasatinib; cisplatin; paclitaxel; vincristine
• Language: croatian
• URN:NBN: urn:nbn:hr:217:550521
• 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: 2017-04-20 12:12:02