Charakterisierung von extrazellulären Vesikeln aus menschlichen Pankreaskrebszellen: die Rolle von p53 auf die Zusammensetzung von PDAC-EVs und deren Einfluss auf NK-Zellen
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Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers, characterized by
its aggressive nature and poor prognosis. A key component in the progression of PDAC is the
tumor microenvironment (TME), where extracellular vesicles (EVs) play a crucial role in
promoting tumor growth, metastasis, and immune evasion. Given the high prevalence of TP53
mutations in PDAC cases and the known influence of p53 on EV biogenesis, this thesis
investigated how p53 mutations affect the composition and function of PDAC-derived EVs,
particularly in their interaction with natural killer (NK) cells, which are critical for tumor
surveillance and suppression. Additionally, two different EV isolation methods were
compared.
This study utilized the human pancreatic cancer cell line Panc-1, carrying the p53R273H
mutation. A stable p53 knockout (p53KO) Panc-1 cell line was generated using CRISPR/Cas9.
EVs were successfully isolated from these cell lines using differential ultracentrifugation (dUC)
and a combination of dUC with size exclusion chromatography (SEC). The impact of p53
mutations and EV isolation techniques on EV size, secretion rate, and EV marker expression
were analyzed through nano-flow cytometry and Western Blot. Furthermore, NK cells were
treated with EVs from Panc-1 and Panc-1-p53KO cells, and their effects on NK cell activation
and cytotoxicity were assessed using flow cytometry and NK cell-killing assays.
The results showed that there were no significant differences between
p53KO-EVs and Panc-1-EVs in terms of particle number, size distribution, EV marker expression,
or immunosuppressive effects on NK cells. EVs from both Panc-1 and Panc-1-p53KO cells
exhibited immunosuppressive effects on NK cell activity. The combination of dUC and SEC
demonstrated that it can lead to a purer EV population, though with lower yield. However,
these EVs appeared to exert a weaker suppressive effect on NK cells compared to EVs isolated
exclusively by dUC. In conclusion, this study highlights that the role of p53 in modulating
PDAC EV secretion and the immunosuppression of NK cells still requires further investigation.
Additional studies on p53 variants, EV cargo, and isolation methods are necessary to better
understand the mechanisms driving PDAC progression and immune evasion.
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Issued: 2026-04-16
Faculty
FB20:Medizin
Language
de
Keywords
PDACEVsNK cellsPankreaskrebsp53
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Magomedov, Arslan: Charakterisierung von extrazellulären Vesikeln aus menschlichen Pankreaskrebszellen: die Rolle von p53 auf die Zusammensetzung von PDAC-EVs und deren Einfluss auf NK-Zellen. : 2026-04-16. DOI: https://doi.org/10.17192/openumr/721.
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This item has been published with the following license: In Copyright