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Journal: Redox Biology
Article Title: Unravelling the anti-cancer mechanisms elicited by non-covalent thioredoxin reductase inhibitors for triple negative breast cancer therapy
doi: 10.1016/j.redox.2025.103980
Figure Lengend Snippet: TXNRD(i)s inhibit TXNRD1 and TXNRD2 enzymes and both enzymes are critical for growth of TNBC cells. A. HCC1806 cells were pretreated with 8VP101 or vehicle control for 1 h, then treated with 10 μM of TRFS-green for 4 h, followed by incubation with MitoTracker-red. Live cells were imaged on a Nikon Ti2E inverted microscope at 20x. Representative images shown. Cytosolic and mitochondrial TXNRD activity on average intensity per cell per field basis was quantified for 4 μM 8VP101 treatment. Average intensity was calculated from n = 3 biological replicates and n = 9 technical replicates. ∗∗∗∗p ≤ 0.0001. B. MDA-MB-231, HCC1806 and MCF-10A cells were transfected with siNeg, siTXNRD1, siTXNRD2, or both siTXNRD1 and siTXNRD2, 10 nM each, and monitored for cell growth every 12 h over 120 h using BioTek BioSpa. Data is shown as fold change normalized to the initial timepoint. Growth curves are averages −/+ sem from n = 3 biological replicates. End-point statistical significance was determined. ∗∗p ≤ 0.005; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001. C. MDA-MB-231 cells were treated with DMSO, 8VP101 or 9VP19 at 10 μM each and monitored for cells growth as in (B). End-point statistical significance was determined. ∗∗∗∗p ≤ 0.0001. D. Cell cycle analysis using PI by flow cytometry of MDA-MB-231 cells. Cells were transfected with 10 nM siTXNRD1, 10 nM siTXNRD2, or a combination of 10 nM siTXNRD1 and 10 nM siTXNRD2 for 24 h. Statistical significance was determined by Kruskal-Wallis test followed by Dunns test comparing siTXNRD1, siTXNRD2, and both siTXNRD1 and siTXNRD2 to siNeg control. Data is presented as the mean±sem. ∗p ≤ 0.05; ns: not significant.
Article Snippet:
Techniques: Control, Incubation, Inverted Microscopy, Activity Assay, Transfection, Cell Cycle Assay, Flow Cytometry