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

Figure 13. Representative Western blot showing FN3K expression in
BT474 cells treated with different compounds. Protein lysates from Figure 15. Representative Western blot showing FN3K expression in
BT474 cells were separated by SDS-PAGE and immunoblotted with an Vero cells after treatment with selected compounds. Vero cell lysates
anti-FN3K antibody. Beta actin served as the internal loading control. were subjected to SDS-PAGE and immunoblotting using an anti-FN3K
A molecular weight marker (lane M) is included for size reference antibody. Beta actin served as a loading control. A molecular weight
(29 kDa and 45 kDa) marker (lane M) is included for size reference (29 kDa and 45 kDa)
Abbreviations: FN3K: Fructosamine-3-kinase
Abbreviations: FN3K: Fructosamine-3-kinase
3.4.3.2. Nrf2 expression analysis
Nrf2 expression remained largely unchanged across all
treatment groups, with oxaliplatin (0.917), capivasertib
(1.008), and lansoprazole (0.964) displaying values
close to those of the untreated control (1.049;
Figure 14). This indicates that these compounds do
not significantly downregulate Nrf2 in BT-474 cells.
The stability of Nrf2 levels suggests the presence of
a resistance mechanism in this cell line that prevents
oxidative stress suppression, distinguishing BT-474
from other models.
These findings underscore the distinct regulatory
responses in BT-474 cells compared to MCF-7 and T-47D,
emphasizing the importance of considering cell-line-
Figure 14. Representative Western blot showing Nrf2 protein expression specific variations in FN3K and Nrf2 modulation when
in BT474 cells after compound treatment. BT474 cell lysates were evaluating therapeutic strategies.
analyzed by SDS-PAGE and probed with an anti-Nrf2 antibody. Beta
actin was used as a loading control. A molecular weight marker (M) 3.4.4. Western blot analysis in Vero cells: Assessing
indicates bands at 29, 45, 67, and 97 kDa specificity and off-target effects
Abbreviations: Nrf2: Nuclear factor erythroid 2-related factor 2
Vero cells, derived from the kidney epithelium of the
African green monkey, serve as a non-malignant control
potentially disrupting metabolic adaptation mechanisms model in comparative studies. Unlike cancer cells, Vero
in BT-474 cells. Lansoprazole (0.802) induced a moderate cells exhibit distinct metabolic and oxidative stress
reduction (~17% decrease), indicating some level of response mechanisms, allowing them suitable for assessing
inhibition, though less pronounced than oxaliplatin. potential off-target effects and the cancer selectivity of
In contrast, capivasertib (0.991) resulted in a negligible tested compounds. Evaluating FN3K and Nrf2 modulation
reduction (~2% decrease), suggesting that it does not in Vero cells following treatment with oxaliplatin,
significantly affect FN3K levels in BT-474 cells. capivasertib, and lansoprazole provides valuable insights

Volume 9 Issue 3 (2025) 215 doi: 10.36922/EJMO025150114