Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: Knockdown, Functional Assay, Quantitative RT-PCR, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Derivative Assay, Co-Culture Assay, Western Blot, Migration

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 depletion in macrophages restrains LUAD growth and metastasis in vivo. (A) Schematic of orthotopic lung implantation and experimental metastasis models using LLC cells mixed with BMDMs expressing shNC or sh-circSMAD4. (B) Representative images of orthotopic lung tumors. (C) Tumor weight of orthotopic implants. (D) Overall survival of mice bearing orthotopic tumors. (E) Immunofluorescence showing F4/80 and circSMAD4 signals in tumor tissues. Scale bar, 50 μm. (F, G) Representative Ki-67 IHC staining and quantification in orthotopic tumors. Scale bar, 50 μm. (H) Representative bioluminescence images of lung tumor burden in the metastasis model. (I) Tumor weight in the metastasis model. (J) Overall survival of mice in the metastasis model. (K–M) Representative IHC staining and quantification of E-cadherin and vimentin in tumors. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: In Vivo, Expressing, Immunofluorescence, Immunohistochemistry

Journal: International Journal of Molecular Medicine

Article Title: 4-Acetylantrocamol LT3 suppresses colorectal cancer growth and metastasis via PI3K/AKT and MAPK pathway modulation

doi: 10.3892/ijmm.2026.5797

Figure Lengend Snippet: LT4 suppresses colony formation, cell migration and EMT in HCT116 cells. (A) Colony formation assay was performed after treating HCT116 cells with various concentrations of LT4 (0.1, 1, 3 and 10 μ M) for 14 days. Colonies were stained with crystal violet and quantification of colony growth was calculated relative to the vehicle-treated Ctrl. (B) Wound healing assay showing inhibition of cell migration by LT4 (0.1, 1, 3 and 10 μ M) at 24, 48 and 72 h post-scratch. Representative images and quantified healing rates are shown. Western blot analysis of EMT-related proteins (C) N-cadherin (140 kDa), (D) E-cadherin (130 kDa) and (E) Vimentin (54 kDa) in HCT116 cells after 24-h LT4 treatment (0.1, 1, 3 and 10 μ M). β-actin (45 kDa) served as the loading control. The corresponding bar graphs show semi-quantification of N-cadherin/β-actin, E-cadherin/β-actin and vimentin/β-actin expressed as fold change relative to the vehicle-treated Ctrl. Data are presented as the mean ± SD from three independent experiments (n=3). * P<0.05, ** P<0.01, *** P<0.001 vs. vehicle-treated Ctrl. LT4, 4-acetylantrocamol LT3; EMT, epithelial-mesenchymal transition; Ctrl, control.

Article Snippet: Most primary antibodies, including those against phosphorylated (p-)AKT (Ser473) (cat. no. 4060), AKT (cat. no. 9272), p-mTOR (Ser2448) (cat. no. 5536), mTOR (cat. no. 2983), p-PI3K p85 (Tyr458) (cat. no. 4228), PI3K p85 (cat. no. 4257), p-ERK1/2 (Thr202/Tyr204) (cat. no. 4370), ERK1/2 (cat. no. 4695), p-p38 (Thr180/Tyr182) (cat. no. 4511), p38 (cat. no. 9212), GSK3β (cat. no. 12456), p-GSK3β (Ser9) (cat. no. 5558), FOXO3a (cat. no. 2497), FOXO1 (cat. no. 2880), p-FOXO1 (Ser256) (cat. no. 9461), p27 kip1 (cat. no. 3686), p21 Cip1/Waf1 (cat. no. 2947), cyclooxygenase-2 (COX-2; cat. no. 12282), Bcl-2 (cat. no. 4223), Bcl-XL (cat. no. 2764), Bax (cat. no. 2772), cytochrome c oxidase subunit IV (COX IV; cat. no. 4850), β-actin (cat. no. 4970), N-cadherin (cat. no. 13116) and E-cadherin (cat. no. 3195) were purchased from Cell Signaling Technology, Inc. [rabbit host; diluted 1:1,000 in 5% BSA (Sigma-Aldrich; Merck KGaA)/PBST (PBS containing 0.1% Tween-20) with 1% NaN 3 ].

Techniques: Migration, Colony Assay, Staining, Wound Healing Assay, Inhibition, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: 4-Acetylantrocamol LT3 suppresses colorectal cancer growth and metastasis via PI3K/AKT and MAPK pathway modulation

doi: 10.3892/ijmm.2026.5797

Figure Lengend Snippet: Proposed schematic model of LT4-mediated anticancer signaling regulation in HCT116 colorectal cancer cells. This schematic summarizes the proposed major pathways and regulatory nodes modulated by LT4, based on transcriptomic, western blotting and molecular docking analyses. LT4 inhibits the PI3K/AKT/mTOR signaling cascade, as reflected by reduced phosphorylation signaling and the annotated functional outcomes (cell viability/cell proliferation and protein synthesis). In addition, LT4 shifts EMT marker expression toward an epithelial phenotype (increased E-cadherin and decreased N-cadherin). In parallel, LT4 modulates the GSK3β-FOXO1/FOXO3a node and differentially regulates MAPK signaling by suppressing ERK phosphorylation while enhancing p38 activation, accompanied by p21 upregulation, consistent with cell-cycle arrest and stress-response phenotypes. LT4 also modulates apoptosis by decreasing the levels of anti-apoptotic proteins (Bcl-2 and Bcl-XL) and increasing Bax expression, alongside mitochondrial destabilization (COX IV) and COX-2 suppression. Key hub genes identified through protein-protein interaction network analysis, SLC3A2, CCND1, PSAT1 and CHAC1, are highlighted as potential mediators linking transcriptomic regulation to these functional outcomes. Black arrows indicate canonical interactions supported by previous studies; red arrows indicate regulatory effects supported by the experimental data in the present study; dashed lines indicate docking-predicted interactions. Arrows indicate activation, whereas blunt-ended lines indicate inhibition. LT4, 4-acetylantrocamol LT3; PI3K, phosphoinositide 3-kinase; AKT, protein kinase B; mTOR, mechanistic target of rapamycin; MAPK, mitogen-activated protein kinase; ERK, extracellular signal-regulated kinase; GSK3β, glycogen synthase kinase 3 β; FOXO, forkhead box O; BAX, COX IV, cytochrome c oxidase subunit IV; COX-2, cyclooxygenase-2; N-cad, N-cadherin; E-cad, E-cadherin; EMT, epithelial-mesenchymal transition; P, phosphorylation; SLC3A2, solute carrier family 3 member 2; CCND1, cyclin D1; PSAT1, phosphoserine aminotransferase 1; CHAC1, ChaC glutathione-specific γ-glutamylcyclotransferase 1.

Article Snippet: Most primary antibodies, including those against phosphorylated (p-)AKT (Ser473) (cat. no. 4060), AKT (cat. no. 9272), p-mTOR (Ser2448) (cat. no. 5536), mTOR (cat. no. 2983), p-PI3K p85 (Tyr458) (cat. no. 4228), PI3K p85 (cat. no. 4257), p-ERK1/2 (Thr202/Tyr204) (cat. no. 4370), ERK1/2 (cat. no. 4695), p-p38 (Thr180/Tyr182) (cat. no. 4511), p38 (cat. no. 9212), GSK3β (cat. no. 12456), p-GSK3β (Ser9) (cat. no. 5558), FOXO3a (cat. no. 2497), FOXO1 (cat. no. 2880), p-FOXO1 (Ser256) (cat. no. 9461), p27 kip1 (cat. no. 3686), p21 Cip1/Waf1 (cat. no. 2947), cyclooxygenase-2 (COX-2; cat. no. 12282), Bcl-2 (cat. no. 4223), Bcl-XL (cat. no. 2764), Bax (cat. no. 2772), cytochrome c oxidase subunit IV (COX IV; cat. no. 4850), β-actin (cat. no. 4970), N-cadherin (cat. no. 13116) and E-cadherin (cat. no. 3195) were purchased from Cell Signaling Technology, Inc. [rabbit host; diluted 1:1,000 in 5% BSA (Sigma-Aldrich; Merck KGaA)/PBST (PBS containing 0.1% Tween-20) with 1% NaN 3 ].

Techniques: Western Blot, Phospho-proteomics, Functional Assay, Marker, Expressing, Activation Assay, Inhibition