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

significant Nrf2 modulation in T-47D cells suggests that In addition, the lack of significant Nrf2 suppression across
FN3K suppression in this model may not directly impact treatments implies that oxidative stress regulatory pathways
oxidative stress signaling, reinforcing the importance in BT-474 cells may be less responsive to these compounds,
of cell-line-specific responses in evaluating potential further reinforcing the importance of considering cell-
therapeutic targets. line-specific variations in therapeutic response.
3.8. Statistical analysis of FN3K and Nrf2 expression 3.9. Selective FN3K inhibition in cancer: Lack of
in BT-474 Cells significant modulation in non-malignant Vero cells
Protein expression changes of FN3K and Nrf2 in Statistical analysis of Western blot data was performed
BT-474 cells were statistically analyzed using Welch’s t-test, using Python-based tools, with appropriate tests applied
with FDR correction applied for multiple testing. Beta actin for normality assessment and pairwise comparisons.
was used for normalization, and statistical significance was Multiple comparison correction was implemented to
defined as p<0.05, as depicted in Figure 19. ensure statistical robustness (Figure 20).
3.8.1. FN3K expression analysis 3.9.1. FN3K expression analysis
Oxaliplatin induced a marginally significant reduction Treatment with oxaliplatin (p=0.846), capivasertib
in FN3K expression (p=0.049), suggesting a moderate (p=0.846), and lansoprazole (p=0.846) did not result in
inhibitory effect on FN3K expression in BT-474 cells statistically significant alterations in FN3K expression
(Figure 19). However, capivasertib (p=0.725) and levels in Vero cells (Figure 20). These results indicate that
lansoprazole (p=0.077) did not significantly alter FN3K none of the tested compounds exert an inhibitory effect on
expression, indicating statistically no significance impact FN3K in this non-malignant cell line, further reinforcing
on this pathway in this cell line. the selective suppression of FN3K in cancerous models.
3.8.2. Nrf2 expression analysis 3.9.2. Nrf2 expression analysis
Similarly, Nrf2 expression remained unaffected across
Nrf2 expression remained unchanged across all treatment all treatment conditions (Figure 20), with oxaliplatin
groups (Figure 19), with oxaliplatin (p=0.300), capivasertib (p=0.600), capivasertib (p=0.624), and lansoprazole
(p=0.739), and lansoprazole (p=0.521) showing no significant (p=0.600) showing no statistically significant changes
differences compared to the untreated control. These results compared to the untreated control. The absence of Nrf2
align with trends observed in T-47D cells, where FN3K modulation suggests that oxidative stress pathways in Vero
suppression did not correlate with notable Nrf2 modulation. cells are not significantly influenced by these compounds.
These findings suggest that while oxaliplatin exerts These findings confirm that FN3K inhibition is
a measurable inhibitory effect on FN3K expression primarily a cancer-specific event, with minimal impact
in BT-474 cells, capivasertib and lansoprazole do not on non-malignant cell lines, such as Vero cells. The lack
significantly impact this metabolic enzyme in this context. of significant FN3K and Nrf2 modulation in these normal
cells supports the hypothesis that FN3K suppression occurs
preferentially in tumor-specific contexts, reinforcing its
potential as a selective therapeutic target in oncology.
3.10. Crosstalk between FN3K and redox-sensitive
pathways
While Nrf2 suppression emerged as a consistent downstream
effect of FN3K inhibition in our study, we recognize that
Nrf2 is a master regulator of cellular redox balance, and its
modulation may impact multiple overlapping pathways.
FN3K has been shown to deglycate lysine and arginine
Figure 20: Western blot densitometric analysis of FN3K and Nrf2 residues on Nrf2, enhancing its nuclear translocation, sMAF
expression in Vero cells. Protein expression levels were normalized binding, and transcriptional activation of ARE-driven genes,
to beta actin. Welch’s t-test with Benjamini-Hochberg correction was such as NQO1, HO-1, and TXNRD1. Our in vitro results
3,10
applied. No significant changes were observed across treatments, as confirmed FN3K and Nrf2 co-downregulation in breast
indicated by the non-significant (ns) labels
Abbreviations: FN3K: Fructosamine-3-kinase; Nrf2: Nuclear factor cancer cells, supporting this mechanistic link. However,
erythroid 2-related factor 2 Nrf2 activity intersects with a wide array of signaling

Volume 9 Issue 3 (2025) 218 doi: 10.36922/EJMO025150114