Karakterizacija adhesoma u stanicama trostruko negativnog karcinoma dojke MDA-MB-231 nakon stabilnog utišavanja gena za integrin alfav

master's thesis

• Title (english): Adhesome characterisation in triple negative breast carcinoma cell line MDA-MB-231 upon stable silencing of integrin subbunit alphav
• Author: Zrinka Vitković
• Mentor: Andreja Ambriović Ristov (mentor)
• Committee member: Marko Močibob (predsjednik povjerenstva)
• Committee member: Biserka Prugovečki (član povjerenstva)
• Committee member: Iva Juranović Cindrić (član povjerenstva)
• Committee member: Morana Dulić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Chemistry); Zagreb
• Defense date and country: 2018-10-29, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry

Abstract

Adhesom integrina je skup proteina koje čine kompleksi adhezije integrina neke stanice. Ovi
kompleksi se nazivaju i fokalne adhezije, a nastaju kada se integrini, transmembranski receptori
sastavljeni od α i β podjedinice, vežu na izvanstanični matriks, što dovodi do nakupljanja
proteina vezanjem na njihovu citoplazmatsku domenu. Cilj je ovog rada bio metodom
spektrometrije masa istražiti adhesom stanica trostruko negativnog karcinoma dojke MDAMB-
231 i usporediti ga s adhesomom stanica klona 231shαv u kojima je, pomoću shRNA
specifične za podjedinicu integrina αv, postignuto stabilno utišavanje te podjedinice. Nakon
izolacije proteina fokalnih adhezija, spektrometrijom masa identificirano je 426 proteina, a
bioinformatičkom analizom utvrđeno je da je njih 107 povezano s fokalnim adhezijama, a 95 s
izvanstaničnim matriksom. Smanjena količina nekih od identificiranih proteina potvrđena je
metodom western blot. U ovome radu definiran je adhesom stanica MDA-MB-231 ovisan o
integrinu αv i doprinjet će otkrivanju mehanizma promjene osjetljivosti ovih stanica na
protutumorske lijekove, cisplatinu i paklitaksel, kao i promjene u sposobnosti migracije i
invazije ovih stanica nakon utišavanja integrina αv.

Abstract (english)

The integrin adhesome is a collection of proteins composed of integrin adhesion complexes in
a given cell. These complexes are also known as focal adhesions which are formed when
integrins, transmembrane receptors comprising an α and β subunit, bind the extracellular matrix
and lead to accumulation of mutimolecular complex of proteins through binding to the
cytoplasmic domains of integrins. The aim of this project was to explore by mass spectrometry,
the adhesome of triple negative breast cancer cells MDA-MB-231 and compare it to the
adhesome of 231shαv cells in which the αv integrin subunit has been silenced by the use of αv
specific shRNA. After focal adhesion isolation, 426 proteins were identified using mass
spectrometry, 107 of which were found to be related to focal adhesions, while 95 of them were
found to be related to the extracellular matrix. Western blots were used to validate the
expression of some of the identified proteins. This project has identified the adhesome of MDAMB-
231 cells and will contribute to research of the mechanism by which the sensitivity of these
cells to cisplatin and paclitaxel is modulated, as well as their ability to modulate migration and
invasion upon aα integrin knockdown.

• Keywords: fokalne adhezije; adhesom; integrin; karcinom
• Keywords (english): focal adhesions; adhesome; integrin; cancer
• Language: croatian
• URN:NBN: urn:nbn:hr:217:784433
• Study programme: Title: Graduate university studies in chemistry; specializations in: Research programme; Course: Research programme; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2020-10-29
• Created on: 2018-11-27 09:15:53