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

these, oxaliplatin exhibited the most potent suppression,
reducing FN3K levels by approximately 66% relative to
the untreated control, closely followed by capivasertib.
These findings suggest that FN3K inhibition may be a
key mechanism underlying the anti-cancer activity of
oxaliplatin and capivasertib. Moderate suppression of
FN3K expression was observed with 1-DMF (0.636) and
ritonavir (0.834), indicating a less pronounced but still
notable inhibitory effect. This suggests that while these
compounds interfere with FN3K function, their effects
are weaker compared to oxaliplatin and capivasertib. In
contrast, amiloride (0.976) had minimal impact on FN3K
expression, implying that its mechanism of action does
not involve FN3K inhibition and may instead act through
alternative metabolic or signaling pathways.
3.4.1.2. Nrf2 expression analysis
Nrf2 expression exhibited the most significant reduction
Figure 9. Representative Western blot showing FN3K protein expression following treatment with amiloride (0.393), representing
following treatment with various compounds. Cell lysates were subjected nearly a 60% decrease relative to the untreated control
to SDS-PAGE and probed with anti-FN3K antibody. Beta actin was used
as a loading control. A molecular weight marker (lane M) is included for (Figure 10). This drastic downregulation suggests that
size reference (29 kDa and 45 kDa) amiloride may interfere with oxidative stress-related
Abbreviations: 1-DMF: 1-deoxy-1-morpholino-D-fructose; FN3K: pathways, potentially sensitizing MCF-7 cells to oxidative
Fructosamine-3-kinase stress and impairing adaptive responses that promote
chemoresistance.
Moderate reductions in Nrf2 expression were observed
with 1-DMF (0.639), ritonavir (0.584), and lansoprazole
(0.683). The suppression induced by lansoprazole (~30%
reduction) aligns with previous studies suggesting that
proton pump inhibitors can modulate oxidative stress and
apoptosis-related mechanisms. In contrast, oxaliplatin
(0.818) and capivasertib (0.812) did not significantly suppress
Nrf2 expression, with levels remaining close to those of the
untreated control. This suggests that while these compounds
effectively inhibit FN3K, they do not substantially impair
oxidative stress defenses, indicating a more selective
mechanism of action. The observed variations in FN3K
and Nrf2 expression across different treatment conditions
provide valuable insights into the mechanistic pathways
Figure 10. Representative Western blot showing Nrf2 protein expression influenced by these compounds. The differential effects on
under different treatment conditions. Cell lysates were analyzed by FN3K and Nrf2 suggest potential therapeutic implications,
SDS-PAGE and immunoblotted with an anti-Nrf2 antibody. Beta actin with some compounds acting primarily as FN3K inhibitors,
was used as a loading control. A molecular weight marker (lane M)
was included for reference, indicating band positions at 29, 45, 67, and while others modulating oxidative stress responses, both of
97 kDa which may contribute to their overall anti-cancer potential.
Abbreviations: 1-DMF: 1-deoxy-1-morpholino-D-fructose; Nrf2:
Nuclear factor erythroid 2-related factor 2 3.4.2. Western blot analysis of FN3K and Nrf2
expression in T-47D Cells
3.4.1.1. FN3K expression analysis Western blot analysis was carried out to quantify FN3K and
FN3K expression was markedly reduced by oxaliplatin Nrf2 protein expression in T-47D cells following treatment
(0.322), capivasertib (0.317), and lansoprazole (0.457), with the selected compounds. The result revealed distinct
indicating a strong inhibitory effect (Figure 9). Among expression patterns of FN3K and Nrf2 across the different

Volume 9 Issue 3 (2025) 213 doi: 10.36922/EJMO025150114