Journal: Oncology Letters

Article Title: Inhibition of apoptosis in breast cancer cells by si-FoxO3a through the protein digestion and absorption signaling pathway

doi: 10.3892/ol.2025.15345

Figure Lengend Snippet: Circos plot of DEPs KEGG pathway enrichment, interaction diagram of DEPs with FoxO3a and western blot results. (A) Circos plot of MCF-7 cell DEPs. (B) Circos plot of MDA-MB-231 cell DEPs. map04974 represents the protein digestion and absorption signal pathway. (C) Protein-protein interaction diagram of DEPs with FoxO3a. (D) The expression of DEPs in MCF-7 cells was assessed using western blotting. (E) The expression of DEPs in MDA-MB-231 cells was verified using western blotting. DEP, differentially expressed protein; KEGG, Kyoto Encyclopedia of Genes and Genomes; si, small interfering RNA; NC, negative control; TPBG, trophoblastic glycoprotein; SLC38A2, solute carrier family 38 member 2; ZYX, zyxin; ASNS, asparagine synthetase.

Article Snippet: A total of 5 μg/lane protein per well, separated by SDS-PAGE (5% concentrating and 5% separating gel), transferred to a PVDF membrane and incubated overnight at 4°C with the following primary antibodies: GAPDH (1:1,000; cat. no. P04406 ; Cell Signaling Technology, Inc.), FoxO3a (1:1,000; cat. no. O43524 ; Cell Signaling Technology, Inc.), trophoblast glycoprotein (TPBG; 1:1,000; cat. no. Q13641 ; Cell Signaling Technology, Inc.), sodium-coupled neutral amino acid symporter 2 (SLC38A2; 1:1,000; cat. no. DF4673; Affinity Biosciences), asparagine synthetase (ASNS; 1:1,000; cat. no. P08243 ; Cell Signaling Technology, Inc.), phosphorylated (p)-FOXO3a (1:1,000; cat. no. O43524 ; Cell Signaling Technology, Inc.) and zyxin (ZYX; 1:1,000; cat. no. Q15942 ; Cell Signaling Technology, Inc.).

Techniques: Western Blot, Expressing, Small Interfering RNA, Negative Control

Journal: Oncology Letters

Article Title: Inhibition of apoptosis in breast cancer cells by si-FoxO3a through the protein digestion and absorption signaling pathway

doi: 10.3892/ol.2025.15345

Figure Lengend Snippet: FoxO3a regulates the protein digestion and absorption signaling pathway. Antitumor drugs act on breast cancer cells, inducing mitochondrial damage and triggering the internal apoptotic program of cancer cells. si-FoxO3a impedes the mitochondrial damage process and affects Gln transport into the cell by decreasing the expression of the amino acid transport protein SLC38A2. This leads to a decrease in the conversion of Gln to glutamate, a decrease in the tricarboxylic acid cycle intermediate product OAA and the catalytic function of ASNS cannot be effectively performed. α-KG deficiency decreases mTOR expression. FoxO3a suppresses mTOR expression, thereby inhibiting ZYX expression and promoting cancer cell apoptosis. FoxO3 also affects TPBG levels through regulation of the Wnt signaling pathway to promote apoptosis in cancer cells. si, small interfering; SLC38A2, solute carrier family 38 member 2; ASNS, asparagine synthetase; α-KG, α-ketoglutarate; mTOR, mechanistic target of rapamycin kinase; TPBG, trophoblastic glycoprotein; ZYX, zyxin; Gluo, glutamate dehydrogenase; OAA, oxaloacetic acid; GS, glutamine synthetase.

Article Snippet: A total of 5 μg/lane protein per well, separated by SDS-PAGE (5% concentrating and 5% separating gel), transferred to a PVDF membrane and incubated overnight at 4°C with the following primary antibodies: GAPDH (1:1,000; cat. no. P04406 ; Cell Signaling Technology, Inc.), FoxO3a (1:1,000; cat. no. O43524 ; Cell Signaling Technology, Inc.), trophoblast glycoprotein (TPBG; 1:1,000; cat. no. Q13641 ; Cell Signaling Technology, Inc.), sodium-coupled neutral amino acid symporter 2 (SLC38A2; 1:1,000; cat. no. DF4673; Affinity Biosciences), asparagine synthetase (ASNS; 1:1,000; cat. no. P08243 ; Cell Signaling Technology, Inc.), phosphorylated (p)-FOXO3a (1:1,000; cat. no. O43524 ; Cell Signaling Technology, Inc.) and zyxin (ZYX; 1:1,000; cat. no. Q15942 ; Cell Signaling Technology, Inc.).

Techniques: Expressing

Journal: Journal of nanobiotechnology

Article Title: Synphilin-1 regulates mechanotransduction in rigidity sensing through interaction with zyxin.

doi: 10.1186/s12951-025-03429-4

Figure Lengend Snippet: Fig. 6 Synphilin-1 regulated substrate rigidity sensing by interacting with zyxin and affecting YAP localization. Under conditions of synphilin-1 overex pression, the following observations were made. Synphilin-1 interacts with endogenous zyxin in the cytosol. This interaction reduces the transportation of zyxin into complexes containing focal adhesion and causes interference in mechanotransduction signaling, especially signaling through the nuclear translocation of YAP. Therefore, synphilin-1-overexpressing cells on the pillar array are unable to spread to further pillars, and they retract their protrusions. In addition, the synphilin-1-overexpressing cells do not respond to substrates of various stiffness through changes in morphology, including spreading area and focal adhesion. These phenomena can be reversed by synphilin-1 knockdown in cells overexpressing synphilin-1

Article Snippet: Each sample was incubated with an anti-zyxin antibody (sc-293448; Santa Cruz Biotechnology) or anti-synphilin-1 antibody (sc-365741; Santa Cruz Biotechnology) overnight at 4 °C.

Techniques: Translocation Assay, Knockdown