Interakcija akvaporina 3 s putem prijenosa signala EGFR/PI3K/Akt u staničnim linijama tumora dojke

master's thesis

• Title (english): Interaction of aquaporin 3 and EGFR/PI3K/Akt signaling pathway in breast cancer cell lines
• Author: Ivan Lučić
• Mentor: Ana Čipak Gašparović (mentor)
• Mentor: Nada Oršolić (komentor)
• Committee member: Nada Oršolić (predsjednik povjerenstva)
• Committee member: Biljana Balen (član povjerenstva)
• Committee member: Duje Lisičić (član povjerenstva)
• Committee member: Tomislav Ivanković (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2022-11-09, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Akvaporin 3 (AQP3), membranska pora, prekomjerno je ispoljena u brojnim tumorima,
uključujući i tumore dojke. Cilj rada bio je istražiti međusobnu povezanost proteina AQP3 i
puta prijenosa signala EGFR/PI3K/Akt praćenjem aktivacije puta preko omjera proteina
pAkt/Akt i utišavanjem proteina AQP3 u staničnim linijama tumora dojke (pozitivne na
receptore za estrogen i progesteron - MCF7, pozitivne na receptor HER2 - SKBR3 i trostruko
negativnu - SUM159) te na ne-tumorskoj liniji epitela dojke (MCF10A). Također, istražen je
utjecaj lokalizacije receptora za epidermalni faktor rasta (EGFR) u lipidne splavi na aktivaciju
ovog puta prijenosa signala. Proučavana je stanična vijabilnost i ekspresija proteina PI3K, Akt,
pAkt i AQP3. Kod stanične linije MCF7 tretman faktorom rasta EGF aktivira protein Akt, ali
ne i protein PI3K. Kod stanične linije SKBR3 disrupcija lipidnih splavi aktivira protein Akt,
neovisno o utišavanju gena za protein AQP3. Kod stanične linije SUM159 dolazi od pojačane
ekspresije proteina PI3K uz tretman faktorom rasta EGF što se gubi utišavanjem gena za
protein AQP3. Kod stanične linije MCF10A tretmani ne utječu na ekspresiju proteina PI3K i
aktivaciju proteina Akt, međutim utišavanjem gena za protein AQP3 raste aktivacija proteina
Akt. Na kraju, u staničnoj liniji SUM159 uočena je ovisnost ekspresije proteina AQP3 o
aktivaciji proteina Akt, ali ne i proteina PI3K.

Abstract (english)

Aquaporin 3 (AQP3), a membrane pore, is overexpressed in numerous tumors, including breast
tumors. The aim of this work was to investigate the relationship between AQP3 and
EGFR/PI3K/Akt signaling pathway by monitoring the activation of the pathway via the
pAkt/Akt ratio and AQP3 silencing in three breast tumor cell lines (positive for estrogen and
progesterone receptors - MCF7, positive for HER2 receptor - SKBR3 and triple negative -
SUM159) and in a non-tumor line of breast epithelium. In addition, the influence of epidermal
growth factor receptor (EGFR) localization in the lipid raft on the activation of this pathway
was investigated. Cell viability and protein expression of PI3K, Akt, pAkt and AQP3 were
investigated. In the MCF7 cell line, treatment with the EGF activates the Akt but not the PI3K.
In the SKBR3 cell line, disruption of lipid rafts activates Akt, independent of gene silencing
for AQP3. In the SUM159 cell line, treatment with EGF results in increased expression of
PI3K, which is lost by silencing the gene for AQP3. In the MCF10A cell line, the treatments
have no effect on PI3K expression and Akt activation, but silencing the gene for the AQP3
increases Akt activation. Finally, in the SUM159 cell line, the expression of the AQP3 was
dependent on the activation of the Akt, but not on PI3K.

• Keywords: AQP3; put prijenosa signala PI3K; lipidne splavi
• Language: croatian
• URN:NBN: urn:nbn:hr:217:098725
• Study programme: Title: Experimental Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2023-09-11
• Created on: 2022-11-22 11:33:55