PhD Scientific Days 2024

Poster Session R - Pharmaceutical Sciences and Health Technologies 2.

Copper Related Protein Expression in KRAS Mutant Cell Lines

Előadó neve

Dr. Üveges, Elina

Neptun code

GH8E3L

Előadó munkahelye

Eötvös Loránd University, Department of Analytical Chemistry

Előadó telefonszáma

+36305626354

Előadó e-mail címe

uvegeselina@gmail.com

Az előadás címe

Copper Related Protein Expression in KRAS Mutant Cell Lines

Szerző(k) neve és munkahelye

Üveges Elina¹

1: Semmelweis Egyetem

Bemutatás módja

Poszter

Szekció

Poster Session R - Pharmaceutical Sciences and Health Technologies 2.

Language of the presentation

English

Preferred session

Pharmaceutical Sciences and Health Technologies

Összefoglaló szövege

Introduction: KRAS mutations are intensively studied nowadays for their importance in human colorectal, lung, and pancreatic cancer. However, these cell types are challenging to target therapeutically. Finding differences in metabolism between wild type and mutant cell lines could be crucial in developing specific therapy for tumors harboring certain KRAS mutations.

Aims: Our earlier studies showed significant differences in copper levels, after long-term copper treatment in KRAS mutant cells, compared to wild type cells. Our aims were to examine the differences in copper related protein expression in response to copper treatment between wild type and KRAS mutant cells.

Methods: We treated SW48 wild type and SW48 G12C, SW48 G12D, and SW48 G12V mutant cells with small concentrations (5 μM) of copper for 72 hours, in the same manner as in our previous long-term copper uptake experiments. After treatment we examined ATP7A (a copper exporter) expression with Western Blot technique. After quantification, we examined the localization of three copper-related proteins: ATP7A, Ctr1 (copper importer) and metallothionein (copper storage protein) with immunofluorescence microscopy.

Results: Western blot experiments have shown high ATP7A expression in SW48 G12V cells, yet the expression remained low in SW48 G12C and SW48 G12D cells, despite similar copper levels after long-term copper treatment in the three mutant cell lines. The immunofluorescent microscopy images show that ATP7A remains in the cytoplasm and is not translocated into the cell membrane after low concentration copper treatment. There is no difference in localization of the three proteins within the cells, comparing the four different cell lines.

Conclusion: There’s a significant difference in long-term copper uptake between SW48 wild type and SW48 KRAS mutant cells. The role of ATP7A is confirmed in the case of SW48 G12V cells, where higher expression of this protein is observed. However, the cause of different copper uptake mechanisms is yet to be explained in the case of SW48 G12C and SW48 G12D cells.

Funding: This work was funded by the Semmelweis 250+ excellency scholarship.

University

Semmelweis University

Supervisor

Dr. Szoboszlai Norbert

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

6883

Start

17:00

End

17:03