Page 225 - EJMO-9-3

Eurasian Journal of
Medicine and Oncology FN3K–Nrf2 axis inhibition in breast cancer

reinforce FN3K as a promising therapeutic target, with reducing Nrf2 expression. Meanwhile, although 1-DMF,
minimal impact on oxidative stress regulatory pathways. amiloride, and ritonavir did not significantly impact FN3K
levels, their ability to suppress Nrf2 suggests possible
3.6. Statistical analysis of FN3K and Nrf2 expression alternative mechanisms of action related to oxidative stress
in MCF-7 Cells and redox balance in MCF-7 cells.
Statistical evaluation of FN3K and Nrf2 protein expression
across various treatment groups was performed using Welch’s 3.7. Statistical analysis of FN3K and Nrf2 expression
t-test, with multiple testing corrections applied through the in T-47D Cells
Benjamini-Hochberg method to control the FDR. Protein In T-47D cells, statistical analysis of FN3K and Nrf2 protein
levels were normalized against beta actin, and all statistical expression was conducted using Welch’s t-test, with FDR
analyses were carried out using the SciPy and statsmodels correction applied for multiple comparisons. Protein levels
libraries in Python. The resulting statistical metrics and were normalized to beta actin, and a p-value below 0.05 was
significance thresholds are illustrated in Figure 17. considered statistically significant, as presented in Figure 18.
3.6.1. FN3K expression analysis 3.7.1. FN3K expression analysis
Oxaliplatin, capivasertib, and lansoprazole significantly Oxaliplatin significantly downregulated FN3K expression
downregulated FN3K expression (p<0.05), indicating (p=0.048), suggesting an inhibitory effect on this metabolic
a strong inhibitory effect on this metabolic enzyme enzyme in T-47D cells (Figure 18). In contrast, capivasertib
(Figure 17). In contrast, 1-DMF (p=0.060), amiloride (p=0.864) and lansoprazole (p=0.072) did not induce
(p=0.678), and ritonavir (p=0.341) did not significantly statistically significant FN3K suppression, indicating minimal
alter FN3K expression, suggesting minimal or no impact or inconsistent effects on FN3K expression in this cell line.
on this pathway in MCF-7 cells.
3.7.2. Nrf2 expression analysis
3.6.2. Nrf2 expression analysis
Nrf2 expression remained largely unaffected across all
All tested compounds, including 1-DMF, amiloride, treatment groups (Figure 18), with oxaliplatin (p=0.300),
ritonavir, oxaliplatin, capivasertib, and lansoprazole, capivasertib (p=0.657), and lansoprazole (p=0.552)
significantly suppressed Nrf2 expression (p<0.05; Figure 17). showing no statistically significant changes compared to
This suggests a potential mechanistic link between FN3K the untreated control. This suggests that, in T-47D cells,
inhibition and oxidative stress modulation, reinforcing the these compounds do not strongly influence oxidative stress
idea that these compounds may influence tumor progression regulatory pathways.
through metabolic and redox regulatory pathways. These findings highlight a differential regulatory
These findings highlight that oxaliplatin, capivasertib, response in T-47D cells compared to MCF-7, where FN3K
and lansoprazole effectively target FN3K while also inhibition was more pronounced and Nrf2 suppression
was significant across multiple treatments. The lack of

Figure 18. Western blot densitometric analysis of FN3K and Nrf2 Figure 19. Western blot densitometric analysis of FN3K and Nrf2
expression in T-47D cells. Protein expression levels were normalized expression in BT-474 cells. Protein expression levels were normalized
to beta actin. Welch’s t-test with Benjamini-Hochberg correction was to beta actin. Welch’s t-test with Benjamini-Hochberg correction was
applied. Significant changes (p<0.05) are marked with an asterisk (*), applied. Significant changes (p<0.05) are marked with an asterisk (*),
while non-significant changes are labeled as ns while non-significant changes are labeled as ns
Abbreviations: FN3K: Fructosamine-3-kinase; Nrf2: Nuclear factor Abbreviations: FN3K: Fructosamine-3-kinase; Nrf2: Nuclear factor
erythroid 2-related factor 2 erythroid 2-related factor 2

Volume 9 Issue 3 (2025) 217 doi: 10.36922/EJMO025150114